Composition of Salt Solutions and Culture Media978-4-431-67875...Composition of Salt Solutions and...

Appendix 1

Composition of Salt Solutions and Culture Media (Listed in Alphabetical Order)

A Salt Solutions

Balanced salt solution (BSSO) for oyster (Tripp et al., 1966) The concentrations are given as g/l,OOO ml solution. NaCI23.50, KCI 0.67, CaClz 1.10, MgClz 2.03, MgS04 2.94, NaHC03 0.02, KzHP04 0.19, glucose 0.50, trehalose 0.50, and phenol red 0.05, made up to 1,000 ml with distilled water.

Buffered physiological solution with antibiotics (BPSA; Hansen, 1979) Unless otherwise indicated, the concentrations are given as g/l,OOO ml solution. NaCI 2.2, KCI 0.12, NazHP04 0.06, glucose 0.8, trehalose 0.8, penicillin 1,000,000 IV, streptomycin 1.0, fungizone 0.0025, and chlortetracycline 0.05, made up to 1,000 ml with distilled water.

Carlson's solution (Carlson et at, 1947) The concentrations are given as g/l,OOO ml solution. NaCI7.0, KCI 0.2, CaClz·HzO 0.2, MgClz·6HzO 0.1, NaHzP04 0.2, NaHC03 0.12, and glucose 8.0, made up to 1,000 ml with distilled water.

Chernin's balanced salt solution (BSS; Chernin, 1963) Unless otherwise indicated, the concentrations are given as g/l,OOO ml solution. NaCI 2.80, KCI 0.15, Na2HP04 0.07, MgS04·7H20 0.45, CaCI2·2H20 0.53, NaHC03 0.05, glucose 1.00, trehalose 1.00, penicillin G 100,000 IV, streptomycin sulfate 0.1, and phenol red (0.4%) 5 ml, made up to 1,000 ml with distilled water.

Chiarandini's saline solution (Chiarandini, 1964) The concentrations are given as g/l,OOO ml solution. NaCI6.45, KCI 0.36, CaCI2·2HzO 0.94, MgS04·7HzO 0.86, and NaHC03 1.54, made up to 1,000 ml with distilled water.

Crab saline (Cooke et at, 1989) NaCl 440 mM, KCI 11.3 mM, CaClz 13.3 mM, MgClz 26 mM, NaZS04 23 mM, HEPES 10 mM, pH adjusted to 7.4 with NaOH. A mixture of antibiotics (penicillin 100 IV/ml, streptomycin 0.1 mg/ml, and fungizone 0.25 g/ml) is added to the Crab saline.

FPl Concentrated complex supplement (Domart-Coulon et al., 1994) Insulin 500 Ilg/ml, hydrocortisone 10-3 M, epidermal growth factor 250 ng/ml, platelet-derived growth factor 100 ng/ml, catalase 500 Ilg/ml, Ex-Cyte VLE (Pentex-Miles) 1 %, egg yolk extract 10%, prostaglandin Fz 10-7 M.

402 Appendix

Gey's salt solution (Gey, 1949) The concentrations are given as gll,OOO ml solution. NaCI 8.0, KCI 0.38, CaCl2 0.13, MgClz-6H20 0.21, Na2HP04·12H20 0.3, KH2P04 0.025, NaHC03 1.0, and glucose 2.2, made up to 1,000 ml with distilled water.

Hydra culture solution (Loomis and Lenhoff, 1956) Chemically defined solution: CaCl2 1O-3M, NaHC03 1O-3M, Convenient solution: NaHC03 20 g, disodium ethylendiaminetetraacetic acid 10 g/ tap water 1,000 ml.

Insect Ringer's solutions (Yamasaki and Narahashi, 1963)

Cockroach Silkworm Japanese beetle

NaCI (g/l,OOO ml) 12.2 8.6

KCI (gl1,000 ml) 0.21 0.10

CaCI, (gil ,000 ml) 0.20 0.33

Buffer * ** *M/15 Na,HPO.: MIlS KH,PO. = 9: 1,50 ml, pH = 7.2 **M/15 Na,HPO. : MIlS KH,PO. = 4 : 6, 50 ml, pH = 6.6

Invertebrate Ringer's Solution

NaCI KCI CaCI, (g/l,OOO ml) (gll,OOO ml) (gll,OOO ml)

Grasshopper 9.0 0.2 0.2

Silkworm 11.2 0.24 0.27

Drosophila 7.5 0.35 0.21

Earthworm 4.4 0.1 0.1

Seawater 25.0 2.6 2.3-3.0 crab

Freshwater 12.0 0.4 1.5 crab

NaHC0 3

(gll,OOO ml)

0.2

0.2

0.2

0.2

0.7

0.2

9.8

0.77

0.38

*

MgCI, (gl1,000 ml)

1.6

0.2

Isotonic buffer for Echinococcus (IBE; Dieckmann and Frank, 1988) NaCl121.00 mM, KCI 5.36 mM, MgS04 0.77 mM, MgCl2 0.93 mM, Na2HP04 0.34 mM, KH2P04 0.44 mM, glucose 5.56 mM, ethyleneglycol-bis-(~-aminoethylether) N,N'tetra acetic acid 0.20 mM, N-2-hydroxyethylpiperazine-N'-2-ethane-sulfonic acid 10.00 mM, pH 7.40.

Laverdure's physiological solution (Laverdure, 1971) The concentrations are given as g/I,OOO ml solution. CaCl2 0.6, MgCI2·6H20 1.0, MgS04·7H20 5.5, NaH2P04 1.0, KCI5.5, trehalose or glucose 1.6, made up to 1,000 ml with distilled water. The pH is adjusted to 7.0 with 5.5 % NaHC03•

Leech Ringer's solution (Nicholls and Kuffler, 1965) NaCl115 mM, KCl4 mM, CaCl2 1.8 mM, Tris maleate buffered with NaOH 10 mM, and glucose 11 mM.

Composition of Salt Solutions and Culture Media 403

Locke's salt solution (Locke, 1895) The concentrations are given as gll,OOO ml solution. NaCI 9.0, KCI 0.42, CaCl2 0.24, and NaHC03 0.2, glucose 1.0 made up to 1,000 ml with distilled water.

Modified Alsever's anti-coagulant solution (Lebel et al., 1996) 115 mM glucose, 27 mM sodium citrate, 11.5 mM EDTA, and 3S2 mM NaCI.

Molluscan balanced salt solution (MMBSS; Machii and Wada, 1989) The concentrations are given as gll,OOO ml solution. NaCI26.22, KCI1.0S, MgS04 3.1S, MgCl2 2.20, CaCl2 1.12, NaHC03 0.30, NaH2P04 0.044, and glucose 0.30, made up to 1,000 ml with distilled water.

Necco's physiological solution (Burnett et al., 1968) The concentrations are given as gll,OOO ml solution. NaCI 25.S00, KCI 0.644, CaCl2 0.644, MgCI2·6H20 3.0S0, KH2P04 0.020, Na2HP04 O.OSO, NaHC03 0.200, Na2S04 0.920, sodium pyruvate 1.500, sodium fumarate 1.050, Lglutamine 1.090, and phenol red 0.020. The pH of the solution is adjusted to 7.5. The antibiotics used in the solution are kanamycin 0.300 gIl, streptomycin 0.100 gIl, and penicillin 0.100 gIl.

Nereis balanced salt solution (NBSS; Heacox et al., 1983) To prepare NBSS, add 6.39 g NaCl, 1.00 g NaHC03, 0.34 g KCI, and 0.169 g CaCI2·2H20 to 742.2 ml natural seawater. Dilute the solution with distilled water to attain the proper osmolarity (920 mOsmollkg). Adjust the pH to between 7.5 and 7.7.

Octopus physiological solution (Necco and Martin, 1963) The concentrations are given as g/l,OOO ml solution. NaCl25.S, KCI 0.644, CaCl2 0.644, MgCI2·6H20 3.0S, KH2P04 0.02, Na2HP04 O.OS, NaHC03

0.2, Na2S04 0.92, sodium pyruvate 1.5, sodium fumarate 1.05, L-glutamine 1.09, and phenol red 0.01, made up to 1,000 ml with distilled water (pH 7.5).

Planarian salt solution (Murray, 1928) The concentrations are given as gll,OOO ml solution. NaCI3.0, CaCI2·6H20 4.0, KCI 0.1, KN03 0.1, MgCl2 1.0 g, MgS04 0.2, NaHC03 0.1, and CaHP04·2Hp 0.1, made up to 1,000 ml with distilled water.

Rinaldini's salt solution (Rinaldini, 1959) The concentrations are given as gll,OOO ml solution. NaCIS.O, KCI 0.20, NaH2P04 0.044, NaHC03 1.0, glucose 1.0, and sodium citrate anhydrous O.SS, made up to 1,000 ml with distilled water.

Ringer's salt solution (Ringer, 1886) The concentrations are given as gil ,000 ml solution. NaCI 7.5, CaCl2 0.125, KCI 0.1, and NaHC03 1.0, made up to 1,000 ml with distilled water.

404 Appendix

SA solution (Furuta and Shim ozawa, 1983) Solution A: NaCI 2,486 mg, KCI 496 mg, CaClz 6 mg, and MgClz 152 mg, made up to 500 ml with distilled water. Solution B: fungizone 5 mg, kanamycin 1,000 mg, and streptomycin 1,000 mg, made up to 40 ml with distilled water. Ready-to-use solution: Solution A 50 ml, solution B 2 ml, 0.1 N NaOH 2 ml, phenol red 0.2 mg, and distilled water 44 ml.

Schistosoma salt solution (Weller and Wheeldon, 1982) The concentrations are given as gll,OOO ml solution. CaClz·2HzO 0040, KCI1.0, MgS04·7HzO 0040, NaC13.0, NaHC03 2.5, and NaHzP04 0.01, made up to 1,000 ml with distilled water.

Snail physiological saline solution (Ivic et aI., 1995) NaCI 100 mM, KCI 4 mM, MgClz 3 mM, CaClz 7 mM.

Tyrode's salt solution (Tyrode, 1910) The concentrations are given as gll,OOO ml solution. NaC18.0, KCI 0.2, CaClz 0.2, NaHC03 1.0, NaHzP04·HzO 0.05, MgClz·6HzO 0.1, glucose 1.0 made up to 1,000 ml with distilled water.

van Harreveld's saline (van Harreveld, 1936) The concentrations are given as gll,OOO ml solution. NaCl 12, CaClz 1.5, KCI 004, MgClz 0.25, and NaHC03 0.2, made up to 1,000 ml with distilled water.

WS washing solution (Rinkevich and Rabinovitz, 1993) MgS04·7HzO 12.06 g (49 mM), CaClz 1.25 g (11.3 mM), NaCl 28.64 g (490 mM), solution of 20 mM HEPES (sodium salt N-2-hydroxyethylpiperazine-N' -2-ethenesulphonic acid) 20 ml, and distilled water to 1,000 ml. The pH of the solution is adjusted to 7.3 with 1 N NaOH. The osmolarity is 1,200 mOsmol/kg. The solution should be sterilized by filtration. The sterilized solution can be stored at 4°C until use.

B Culture Media

Barnacle organ culture medium IV (Fyhn and Costlow, 1975) NaCl466 mM, KCl8 mM, CaClz 20 mM, MgClz 12 mM, NaHC03 10 mM, glucose 0.1 %, bovine embryo extract (Difco Laboratories) 1 %, and TC-Yeastolate (Difco Laboratories) 0.1%.

Basch and DiConza's medium C (Basch and DiConza, 1973) The concentrations are given as mgll,OOO ml medium. NaCl1,300, KCI 97, MgClz·6HzO 325, NazHP04 17, CaClz·2HzO 558, NaHC03 1,769, Llysine 5.8, L-histidine 2.0, L-arginine 204, L-aspartic acid 5.6, L-threonine 6.2, L-serine 12.0, L-glutamic acid 904, L-proline 5.8, glycine 4.8, L-alanine 4.8, L-valine 3.2, L-me-


thionine 0.03, L-isoleucine 2.0, L-Ieucine 2.9, L-phenylalanine 1.B, diphosphopyridine nucleotide 7.0, triphosphopyridine nucleotide 2.0, coenzyme A 2.5, cocarboxylase 2.0, flavin-adenine dinucleotide 2.0, biotin 1.0, folic acid 1.0, choline chloride 1.0, nicotinamide 1.0, o-calcium pantothenate 1.0, pyridoxine hydrochloride 1.0, thiamine hydrochloride 1.0, riboflavin 0.1, inositol1.B, cyanocobalamin 10.0, nicotinic acid 2.0,paminobenzoic acid 2.0, pyridoxine hydrochloride 2.0, uridine 2.0, deoxyadenosine 10.0, deoxycytidine 10.0, deoxyguanosine 10.0, thymidine 10.0, cytosine 1.0, glucuronolactone 1.B, glutathione 10.0, phenol red 20.0, and glucose 100.0. The pH of the solution should be adjusted to 7.2-7.4. The osmolarity of the solution is 155 mOsmol/kg. The medium is supplemented with 10% heat-inactivated FBS and 10% whole egg ultrafiltrate. Sometimes hemolymph from adult Biomphalaria glabrata is added at a concentration of 5% after inactivation at 60°C for 10 min. An antibiotic mixture (penicillin G 100 IV, streptomycin sulfate 100 !lg, gentamicin sulfate 10 !lg/ml) is added to the medium.

Botryllid cell culture medium (BCCM; Rinkevich and Rabinovitz, 1993) The medium basically consists of Dulbecco's modified Eagle's medium x5 (DMEM) and a salt solution, the osmolarity of which is adjusted to that of botryllid ascidian hemolymph. The salt solution is made up of two major components at a 2x concentration. Component A: NaCI13.67 g, KCI 0.412 g, and CaClz·2HzO 0.721 g, made up to 125 ml with distilled water. Component B: MgS04·7HzO 5.57 g and MgClz·6HzO 3.05 g, made up to 125 ml with distilled water. Both components are mixed (1 : 1), and the solution is sterilized by filtration through a membrane filter with a 0.2 !lm pore size. Both stock solutions can be stored at 4°C. The ready-to-use BCCM should be prepared before use. Each 100 ml BCCM is made by mixing 20 ml DMEM (x5), 50 ml salt solution, 26 ml distilled water, 2 ml L-glutamine solution (200 mM),2 ml HEPES buffer (1 M,pH 7.0) and 1 ml antibiotic mixture (penicillin sodium salt 104 IU, streptomycin 10 mg, and nystatin 1,250 IU). The medium is sterilized by filtration through a membrane filter with a 0.2 !lm pore size, and can be stored at 4°C for up to 3 weeks. Heat-inaysate (5%) 10.0 ml, and yeast extract (10%) 5.0 ml.

Cecil's medium modified by Heacox et al. for Nereis cell culture (Heacox et al., 1983) Distilled water 147 ml, NaCI 6,390 mg, NaHC03 1,000 mg, KCI 340 mg, CaClz·2HzO 169 mg,glucose 1,000 mg,galactose 500 mg,fucose(o) 250 mg,fucose(L) 250 mg,minimal essential medium (MEM) essential amino acids (lOOx) 20.0 ml, MEM non-essential amino acids (100x) IB.O ml, MEM vitamin (100x) 3.3 ml, glutamine 200 mg, gentamicin 1.0 mg, penicillin 300,000 IV, Nereis coelomic fluid ultrafiltrate 0-20 ml, horse serum 0-100 ml, hens' whole egg ultrafiltrate 0-100 ml, and Nereis vitellin 0 or 500 mg.

Chernin's nutrient medium (Chernin, 1963) Chernin's BSS 850.0 ml, bovine amniotic fluid 110.0 ml, beef embryo extract 12.5 ml, horse serum 12.5 ml, lactalbumin hydrolysate (5%) 10.0 ml, and yeast extract (10%) 5.0 ml.

406 Appendix

Cp medium (Miltenburger et aI., 1984) The concentrations are given as mgll,OOO ml medium. KCI 2,870.0, CaCI2·2H20* 1,320.0, MgCI2·6H20* 2,280.0, MgS04·7H20* 2,780.0, NaH2P04·2H20* 1,140.0, NaHC03 350.0, L-arginine hydrochloride 350.0, L-aspartic acid 175.0, L-asparagine 175.0, L-alanine 112.5, ~-alanine 100.0, L-cystine 11.0, L-glutamic acid 300.0, L-glutamine 300.0, glycine 320.0, L-histidine hydrochloride 1,250.0, L-isoleucine 25.0, L-Ieucine 37.5, L-Iysine hydrochloride 312.5, L-methionine 25.0g, L-proline 175.0, L-phenylalanine 75.0, DL-serine 550.0, L-threonine 87.5, L-tryptophan 50.0, L-tyrosine 25.0, L-valine 50.0, p-aminobenzoic acid 0.010, biotin 0.005, D-calcium pantothenate 0.010, choline chloride 0.100, folic acid 0.010, i-inositol 0.010, nicotinic acid 0.010, pyridoxine hydrochloride 0.010, riboflavin 0.010, thiamine hydrochloride 0.010, glucose 1,000.0, and tryptose 2,600.0, made up to 1,000 ml with distilled water. The medium is supplemented with 10% FBS. The pH is adjusted to 6.4 with 1 N KOH.

Crab integument medium I and II (Ballard et aI., 1993) Stock solutions: Modified medium 199 (MI99). Dissolve 10,900 mg Medium 199 (Sigma) in 900 ml deionized-distilled water (DDW).Add 5,960 mg HEPES,200 mg MgCI2·6H20, 15,000 mg NaCI, and 200 mg L-proline and adjust the pH to 7.5 with 1 N NaOH or 1 N HCI. Make the solution up to a final volume of 1,000 ml with DDW. Sterilize with a 0.22-l1m filter and store at 4°C. Antibiotic cocktails (HCA). In 900 ml DDW, dissolve 100 mg Benomyl (Dragon Chemical), 50 mg gentamicin sulfate, 25 mg nystatin, 100 mg penicillin G. When in solution, add 1.65 ml penicillin-streptomycin solution (Sigma), 0.5 ml gentamicin sulfate solution (Sigma), and 0.5 ml fungizone (Gibco).Adjust the pH to 7.5 with 1 N NaOH or 1 N Hel. Make the solution up to a final volume of 1,000 ml with DDW. Sterilize through a 0.22-l1m filter and store in 7.65 ml aliquots at -20°C (thaw before use in final medium). Antibiotic cocktails (LCA). Sterilize the penicillin-streptomycin solution (Sigma), gentamicin sulfate solution (Sigma), and fungizone (Gibco) by passage through a 0.22-l1m filter. Into separate sterile tubes, aliquot 1.65 ml penicillin-streptomycin, 0.5 ml gentamicin sulfate and 0.5 ml fungizone. Store the aliquots at -20°C (thaw before use in final medium). Complete medium I: mix 95 ml M119 and 5 ml FBS. Take 7.65 ml this mixture and add a 7.65 ml HCA aliquot. Sterilize the mixture by passage through a 0.22-l1m filter and store the solution at 4°C (warm to room temperature before use). Complete medium II: mix 95 ml M119 and 5 ml FBS. Take 2.65 ml this mixture and add one aliquot of LCA. Sterilize the solution by passage through a 0.22-l1m filter and store at 4°C (warm to room temperature before use).

Crayfish integument medium (Daig and Spindler, 1979) Mix 2 parts of van Harrevelt saline with 1 part horse serum; osmolarity 349 mOsmoll kg, pH 7.6.

CSM-2F medium (Mitsuhashi, 1967) Unless otherwise indicated, the concentrations are given as mg/1,000 ml medium. NaH2P04·Hp 500, MgCI2·6H20 1,200, MgS04·7H20 1,600, KCl1,200, CaCI2·2H20 400, glucose 800, fructose 800, lactalbumin hydrolysate 5,200, Bacto-peptone (Difco Laboratories) 5,200, choline chloride 400, TC-Yeas to late 2,000, TC-199 medium 200 ml, FBS


200 ml, and dihydrostreptomycin sulfate 100, made up to 1,000 ml with distilled water. The pH of the medium is adjusted to 6.2 with KOH.

Dulbecco's modified Eagle's medium (Morton, 1970) The concentrations are given as mg/l,OOO ml medium. CaClz 200, Fe(N03)3·9HzO 0.1, KCI 400, MgS04 97.67, NaCI 6,400, NaHC03 3,700, NaHzP04·HzO 125, glucose 1,000, phenol red 15, sodium pyruvate 110, L-arginine hydrochloride 84, L-cystine dihydrochloride 62.57, L-glutamine 584, glycine 30, L-histidine hydrochloride monohydrate 42, L-isoleucine 104.8, L-Ieucine 104.8, L-Iysine hydrochloride 146.2, L-methionine 30, L-phenylalanine 66, L-serine 42, L-threonine 95.2, tryptophan 16, L-tyrosine 2Na·2HzO 103.79, L-valine 93.6, o-calcium pantothenate 4.0, choline chloride 4.0, folic acid 4.0, myoinositol 7.0, niacinamide 4.0, pyridoxal hydrochloride 4.0, riboflavin 0.4, and thiamine hydrochloride 4.0.

Durchon and Schaller's medium for Annelida (Durchon and Schaller, 1963) Mix the following: 7 drops of 1 % agar in sterilized natural seawater containing 0.1 % glucose, 3 drops of sterilized seawater containing 0.1% glucose, 1 drop of 50% egg albumin in sterilized seawater containing 0.1 % glucose, 1 drop of horse serum (Difco Laboratories), 1 drop of 50% chicken embryo extract in sterilized seawater containing 0.1 % glucose, 100 IV of penicillin G, and 0.009 mg of streptomycin.

Gottschewski's medium (Gottschewski, 1960) The concentrations are given as mg/l,OOO ml medium. L-Arginine 170, L-cysteine 40, L-cystine 3, glycine 12, L-histidine 196, L-Ieucine 200, Llysine 140, o-methionine 50, L-phenylalanine 200, L-threonine 120, L-tryptophan 20, glucose 1,600, mannose 1,600, fructose 1,600, choline 5, folic acid 0.5, inositol 1.0, nicotinic acid 5.0, pantothenate 1.0, thiamine 0.5, riboflavin 0.5, pyridoxine 1.0, DPN 10, TPN 5, adenine deoxyribonucleoside 0.5, guanine deoxyribonucleoside 0.5, uridine-5-triphosphate O.5,deoxycytidine diphosphate 0.5,deoxy-5-methylcytidilic acid 0.5, thymidine diphosphate 0.5, ATP 5.0, RNA 50, DNA 50, NaCI 5,000, sodium acetate 50, NazHP04·5HzO 10,000, KC1300, CaClz·2HzO 20, MgClz·6HzO 300, MgS04·7HzO 300, glutathione 500, peptone 1,000, glycylglycine 1,000, casein hydrolysate 5,000, and yeast extract 15. The pH of the medium is adjusted to 7.0-7.2 with 1 N NaOH and 1 N HCl.

Grace's medium (GMA; Grace, 1962) The concentrations are given as mg/l,OOO ml medium. NaHzP04·2HzO 1,140, NaHC03 350, KCI2,240, MgClz·6HzO 2,280, MgS04·7HzO 2,780, CaClz l,OOO, L-a-alanine 225, ~-alanine 200, L-arginine hydrochloride 700, L-asparagine 350, L-aspartic acid 350, L-cystine hydrochloride 25, L-glutamic acid 600, L-glutamine 600, glycine 650, L-histidine 2,500, L-isoleucine 50, L-Ieucine 75, L-Iysine hydrochloride 625, L-methionine 50, L-phenylalanine 150, L-proline 350, oL-serine 1,100, L-threonine 175, L-tryptophan 100, L-tyrosine 50, L-valine 100, sucrose 26,680, glucose 700, fructose 400, malic acid 670, a-ketoglutaric acid 370, succinic acid 60, fumaric acid 55, thiamine hydrochloride 0.02, riboflavin 0.02, calcium pantothenate 0.02, pyridoxine hydrochloride 0.02, p-aminobenzoic acid 0.02, folic acid 0.02, niacin 0.02, isoinositol 0.02, biotin 0.01, choline chloride 0.2, penicillin G sodium salt 30, and streptomycin sulfate 100, made up to 1,000 ml with distilled water. The pH is adjusted to 6.5 with KOH. The medium is used by supplementing it with 3% heat-inactivated hemolymph of Lepidoptera or 10% FBS.

408 Appendix

Haliotis discus culture medium (Naganuma et aI., 1996) Leibovitz's L-15 medium (Gibco) with the following components added: NaCI 1.5% (w/v) final concentration (originally 0.85%), NaHC03 2 mM final concentration, antibiotic-antimycotic solution (lOOx; Gibco; amphotericin B 25 mg/ml, ampicillin 5 mgt ml, and streptomycin sulfate 10 mg/ml), lipid mixture (lOOx; Sigma), and FBS (lOx; GIBCO). The final pH of the culture medium should be 7.8.

Hansen's M-182 medium (Hansen, 1976) The concentrations are given as mg/l,OOO ml culture medium. NaCI2,800, KC1150, CaCl2 530, MgS04·7H20 450, Na2HP04 70, NaHC03 50, L-alanine 6.0, L-arginine 4.2, L-asparagine 2.0, L-aspartic acid 6.0, L-cysteine 6.0, L-cystine 2.4, Lglutamic acid 10.0, L-glutamine 20.0, glycine 6.0, L-histidine 4.0, L-isoleucine 5.2, Lleucine 5.2, L-Iysine 5.8, L-methionine 1.5, L-phenylalanine 3.5, L-proline 6.0, L-serine 12.0, L-threonine 4.8, L-tryptophan 0.8, L-tyrosine 3.6, L-valine 4.7, galactose 2,000, glucose 860, trehalose 860, biotin 0.038, calcium folinate 0.038, o-calcium pantothenate 0.075, cyanocobalamin 0.038, folic acid 0.075, N-acetylglucosamine 0.150, niacin 0.075, niacinamide 0.075, pantethine 0.038, p-aminobenzoic acid 0.075, pyridoxal hydrochloride 0.038, pyridoxamine hydrochloride 0.038, pyridoxine hydrochloride 0.075, riboflavin 0.075, thiamine hydrochloride 0.075, thioctic acid 0.038, Bacto-peptone (Difco Laboratories) 4,500, and FBS 130 ml. The pH of the medium should be 7.0-7.2 and the osmolarity 185 mOsmollkg.

Hansen's M-260 medium (Hansen, 1976) The concentrations are given as mg/l,OOO ml culture medium. NaCI 1,792, KCI 96, CaCl2 339, MgS04·7H20 288, Na2HP04 45, NaHC03 32, L-alanine 3.8, L-arginine 2.7, L-asparagine 1.3, L-aspartic acid 3.8, L-cysteine 3.8, L-cystine 1.5, Lglutamic acid 6.4, L-glutamine 20.0, glycine 3.8, L-histidine 2.6, L-isoleucine 3.3, L-Ieucine 3.3, L-Iysine 3.7, L-methionine 1.0, L-phenylalanine 2.1, L-proline 3.8, L-serine 7.7, L-threonine 3.1, L-tryptophan 0.5, L-tyrosine 2.3, L-valine 3.0, galactose 1,280, glucose 550, trehalose 550, biotin 0.024, calcium folinate 0.024, o-calcium pantothenate 0.048, cyanocobalamin 0.024, folic acid 0.048, N-acetylglucosamine 0.096, niacin 0.048, niacinamide 0.048, pantethine 0.024, p-aminobenzoic acid 0.048, pyridoxal hydrochloride 0.024, pyridoxamine hydrochloride 0.024, pyridoxine hydrochloride 0.048, riboflavin 0.048, thiamine hydrochloride 0.048, thioctic acid 0.024, adenosine-3'(2')phosphoric acid 0.347, cytidine-3'(2')phosphoric acid 0.323,guanosine-3'(2')phosphoric acid 0.363, uridine-3'(2')phosphoric acid 0.324, thymine 0.126,lactalbumin hydrolysate (EdaminType S; Humco-Sheffield) 4,500, and FBS 130 ml. The pH of the medium should be 7.0-7.2 and the osmolarity 154 mOsmollkg.

Hansen's S-301 medium (Hansen, 1976) The concentrations are given as mg/l,OOO ml culture medium. NaCl462 KC1352, CaCl2 132, MgS04·7H20 814, Na2HP04 154, KH2P04 99, NaHC03 88, L-arginine 88, L-aspartic acid 88, ~-alanine 110, L-cysteine 13, L-cystine 22, L-glutamic acid 176, L-glutamine 396, glycine 55, L-histidine 88, L-isoleucine 33, L-Ieucine 33, Llysine 363, L-methionine 176, L-phenylalanine 33, L-proline 374, L-serine 55, L-threonine 77, L-tryptophan 22, L-tyrosine 110, L-valine 66, galactose 1,300, glucose 440, trehalose 440, fumaric acid 22, a-ketoglutaric acid 44, malic acid 22, succinic acid 22, lactalbumin hydrolysate (Edamin-Type S; Humco-Sheffield) 4,500, yeast hydrolysate


440, and FBS 130 m!. The pH of the medium should be 7.0-7.2 and the osmolarity 155 mOsmollkg.

Haliotis rufescens culture medium (Naganuma et al., 1994) Leibovitz's L-15 medium (Gibco) with the following components added: NaCI3% (w/ v) final concentration (originally 0.85%), L-glutamine 2 mM final concentration, NaHC03 2 mM final concentration, antibiotic solution (100x; amphotericin B 25 mgt ml, ampicillin 5 mg/ml, penicillin G 10,000 IU/ml, and streptomycin sulfate 10 mg/ml), lipid mixture (1,OOOx; Sigma), yeast extract 200 mg/ml (100x; Sigma), calf serum (10-20x). The final pH of the solution should be 7.6 and final osmolarity 1,080 mOsmollkg.

Heacox et al?s Medium G (Heacox et aI., 1983) Natural seawater (sterile) 742 ml, distilled water 218 ml, NaCI 6,390 mg, NaHC03

1,000 mg, KCl340 mg, CaClz·2HzO 169 mg, glucose 1,000 mg, galactose 500 mg, fucose (D) 250 mg, fucose (L) 250 mg, gentamicin 1.0 mg, penicillin 300,000 IU, Nereis coelomic fluid ultrafiltrate 0-20 ml, and Nereis vitellin 0 or 500 mg.

Honeybee medium (Kaatz et al., 1985) The concentrations are given as mg/l,OOO ml culture medium. NaCI 1,580, KCI 1,790, CaClz·2HzO 1,029, MgClz·6HzO 406, MgS0 4·7HzO 493, NaHzP04·HzO 690, NaHC03 336, L-a-alanine 300, L-arginine 500, L-asparagine 200, Laspartic acid 100, L-cysteine 50, L-glutamic acid 250, L-glutamine 1,000, glycine 200, Lhistidine 200, L-isoleucine 100, L-Iysine 550, L-methionine 50, L-phenylalanine 100, Lproline 3,300, DL-serine 200, L-threonine 150, L-tryptophan 50, L-tyrosine 100, L-valine 150, glucose 4,000, fructose 2,500, sucrose 55,370, PIPES 7,560, bovine serum albumin 50,000, penicillin G ISO, streptomycin 250. The pH is adjusted to 6.70 with 1 N KOH and 1 N HC!.

Hydra growth medium (Yu-Ying et aI., 1963) Eagle's medium modified by Swin and Parker (1958) 10 m1,horse serum 100 ml,penicillin 500 IU, Fumidil B 200 mg, NaCI 5.8 mg, MgS04·7HzO 100 mg, NaHC03 168 mg, KCI 7.5 mg, CaClz III mg, dextrose 1,000 mg, phenol red 10 mg, streptomycin 12.5 mg, triple-distilled water 800 ml, and 0.15 M phosphate buffer, to maintain the pH of the medium at 6.8, made up to 1,000 ml with distilled water.

IPL-41 medium (Weiss et aI., 1981) The concentrations are given as mg/l,OOO ml culture medium. NaHzP04·HzO 1,160, NaHC03 350, KCl1,200, CaClz 500, MgS04·7HzO 1,880, B-alanine 300, L-arginine hydrochloride 800, L-asparagine 1,300, L-aspartic acid 1,300, L-cystine 100, L-glutamic acid 1,500, L-glutamine 1,000, glycine 200, L-histidine 200, hydroXY-Lproline 800, L-isoleucine 750, L-leucine 250, L-Iysine hydrochloride 700, L-methionine 1,000, L-proline 500, L-phenylalanine 1,000, oL-serine 400, L-threonine 200, L-tryptophan 100, L-tyrosine 250, L-valine 500, sucrose 16,500, glucose 2,500, maltose 1,000, malic acid 53.6, a-ketoglutaric acid 29.6, succinic acid 4.8, fumaric acid 4.4, thiamine hydrochloride 0.08, riboflavin 0.08, calcium pantothenate 0.008, pyridoxine hydrochloride O.4,p-aminobenzoic acid 0.32, folic acid 0.08, niacin 0.16, inositol 0.4, biotin 0.16, cyanocobalamin 0.24, choline chloride 20, ZnClz 0.04, MnCl2·4H20 0.02, CuCl2·2HzO 0.2, (NH4)Mo70Z4·4H20 0.04, CoClz·6HzO 0.05, and FeS04·7H20 0.55. The osmolarity of the medium should be 360-375 mOsmollkg. FBS 10% may be added to the medium.

410 Appendix

K-6 medium (Horikawa, 1959) The concentrations are given as mg/l,OOO ml culture medium. NaCl 7,000, KC1200, CaCI2·2H20 20, MgCI2·6H20 100, NaHC03 50, NaH2P04·2H20 200, glucose 800, casein hydrolysate 50,000, tryptophan 5,000, cystine 1,000, cysteine 260, oL-kynurenine 4 g, oL·3-hydroxykynurenine 2 g, ribonucleic acid 500, thiamine 0.05, riboflavin 0.05, pyridoxine hydrochloride 0.125, niacinamide 0.125, calcium pantothenate 0.05, ascorbic acid 250, biotin 0.05, folic acid 0.05, choline chloride 2.5, inosite 0.25,p-aminobenzoic acid 0.25, vitamin A 0.5, vitamin B12 0.5, sodium acetate SO, glutathione 10, glutamic acid 100, diphosphopyridine nucleotide (DPN) 1.0. The pH of the medium should be adjusted to 7.2 with 1 N KOH.

Kuroda and Tamura's medium for Drosophila melanotic tumors (Kuroda and Tamura, 1955) The concentrations are given as mg/1,OOO ml culture medium. NaCI 7,000, KC1200, CaCI2·2H20 20, MgCI2·6H20100, NaHC03 SO, NaH2P04·2Hp 200, glucose 800, casein hydrolysate 50,000, tryptophan 1,000, cystine 1,000, cysteine 260, ribonucleic acid 500, thiamine 0.05, riboflavin 0,05, pyridoxine hydrochloride 0.125, niacinamide 0.125, calcium pantothenate 0.05, ascorbic acid 250, biotin 0.05, folic acid 0.05, choline chloride 2.5, inosite 0.25, p-aminobenzoic acid 0.25, cyanocobalamin 0.5, vitamin A 0.5, sodium acetate SO, glutathione 10, glutamic acid 100, and DPN 1.0.

L-15 medium (Leibovitz, 1963) The concentrations are given as mg/1 ,000 ml culture medium. NaCI 8,000, KCI 400, MgS04·7H20 200, MgCl2·6H20 200, CaCl2 140, Na2HP04 190, KH2P04 60, oL-a-alanine 450, L-arginine 500, L-asparagine 250, L-cysteine 120, glycine 200, L-histidine 250, DL-isoleucine 250, L-Ieucine 125, L-Iysine 75, DL-methionine 150, DL-phenylalanine 250, L-serine 200, DL-threonine 600, L-tryptophan 20, DL-valine 200, choline 1, folic acid 1, inositol 2, nicotinamide 1, DL-Ca pantothenate 1, pyridoxine hydrochloride 1, sodium riboflavin-5-phosphate 0.1, thiamine monophosphate 1, Lglutamine 300, L-tyrosine 300, sodium pyruvate 550, o{ + )galactose 900, and phenol red 10.

L-15B (Munderloh and Kurtti, 1989) Unless otherwise indicated, the concentrations are given as mg/1 ,000 ml culture medium. Leibovitz's L-15 medium powder (Gibco) 1 package 03.88g), L-aspartic acid 299, Lglutamic acid 500, L-glutamine 292, L-proline 300, a-ketoglutaric acid 299, o-glucose 2,239, mineral stock* 1 ml, vitamin stock** 1 ml. The pH of the medium is adjusted to 6.4-6.6 with 1 N NaOH. *Mineral stock (x103) contains stock solutions A, B, and C. The components of stock solution A (xl0S) are: CoCI·6H20 20 mg, CuS04·5H20 20 mg, MnS04·H20 160 mg, ZnS04·7H20 200 mg, and distilled water 100 ml. The components of stock solution B (xlOS) are NaMo04·2H20 20 mg and distilled water 100 ml. The components of stock solution C (xlOS) are Na2Se03 20 mg and distilled water 100 ml. The complete mineral stock (xl03) are: reduced glutathione 1,000 mg, ascorbic acid 1,000 mg, FeS04·7H20 50 mg, stock solution A 1 ml, stock solution B 1 ml, stock solution C 1 ml, and distilled water to 100 ml. **Vitamin stock (xl03): p-aminobenzoic acid 100 mg, cyanocobalamin 50 mg, o-biotin 10 mg, and distilled water 100 ml.


Laverdure's medium (Laverdure, 1971) Laverdure's medium is made by mixing 1 part of horse serum (Difco Laboratories; dissolve the content of one ampoule in 5 ml bidistilled water), 1 part chicken embryo (11 days old) extract (Difco Laboratories; dissolve the contents of one ampoule in 8 ml Laverdure's physiological solution), 1 part Laverdure's physiological solution containing 2% peptone (Merck, Darmstadt, Germany), and 2 parts Laverdure's physiological solution containing 1 % agar. Penicillin (20,000 IU) and streptomycin (0.5 mg) are added to 20 ml medium. The pH of the medium is adjusted to 6.84-7.1 with 5.5% NaHC03•

Leloup's medium 1 (Leloup, 1964) The concentrations are given as mg/l,OOO ml culture medium. NaCI6,500, KC1250, MgS04·7H20 350, CaCl2 300, trehalose 800, glucose 700, malic acid 500, a-ketoglutaric acid 250, succinic acid 50, fumaric acid 50, yeast extract 1,000, and lactalbumin hydrolysate 10,000. The pH of the medium is adjusted to 7.0-7.1 with 1 N NaOH.

Limulus organ culture medium (Bayer and Barlow, 1978) The concentrations are given as mg/l,OOO ml culture medium. NaCI 16,100, KCI 276, CaCI2·2H20 1,184, MgCI2·6H20 1,423, MgS04·7H20 5,669, medium 199 with Hanks salts (lOx; Gibco) 100 ml, horse serum 100 ml (Gibco), glucose 2,703, L-glutamine 292, retinol* (2 gil, Sigma) 10 ml, tocopherol* (200 mg/l, Sigma) 10 ml,penicillin-streptomycin (10,000 U/ml) 20 ml,HEPES (50 mM,Sigma) 10 ml,and TES (N-tris (Hydroxymethyl) methyl-2-aminoethane-sulfonic acid; 2-( (2Hydroxy-l, I-bis (hydroxymethyl)-ethyl) amino) ethansulfonic acid; 50 mM, Sigma) 10 ml, made up to 1,000 ml with distilled water. The pH should be adjusted to 7.3. * Add as an alcoholic solution.

Medium A-12 (Tripp et aI., 1966) Mix 5% casamino acid 1.0 ml, 10% yeast extract 10 ml, 5% lactalbumin hydrolysate 10 ml, calf serum 10 mI, and BSSO (see Salt Solutions) 969 ml. The pH of the medium should be 7.2-7.4.

Medium 1779 for Schistosoma (Weller and Wheeldon, 1982) Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium. CaCI2·2H20 400, KCll,OOO, MgS04·7H20 400, NaCI3,000, NaHC03 2,500, NaH2P04 10, NaOH 200, HEPES 2,380, ZnS04·7H20 0.022, FeS04·7H20 0.01, MnS04·4H20 0.004, CuS04 0.0005, CoS04·7H20 0.00044, H3B03 0.00011, L-alanine 1,000, DL-cysteine hydrochloride 50, L-glutamine 150, glycine 250, L-histidine 1,000, hydroXY-L-proline 500, L-Iysine hydrochloride 100, L-ornithine hydrochloride 10, L-serine 250, Eagle's BME essential amino acids (Gibco) 1 ml, glutathione 10, L-histidyl-L-alanine 25, glucose 12,000, trehalose 500, adenine 2.5, cytosine 0.5, guanine hydrochloride 1.0, thiamine 0.5, uracil 1.0, xanthine sodium salt 1.0, orotic acid sodium salt 50.0, a-ketoglutaric acid sodium salt 200, DL-carnitine hydrochloride 5.0, choline chloride 20.0, menadione sodium bisulfite/albumin* 1 flg, vitamin B12 10.0, Eagle's BME vitamin block (Gibco) 5.0 ml, dexamethasone 392.5 ng, insulin 5.0, triiodo-L-thyronine 10 flg, Cohn's fraction IV 100, FBS 20.0 ml, gelatin 1,000, yeast extract/serum** 1.0 ml, phenol red 20, serotonin creatinine sulfate 0.4, and cholesterol/albumin***.

412 Appendix

*Menadione sodium bisulfite/albumin: 0.002 g menadion sodium bisulfite is homogenized with 0.24 g bovine albumin in 20 ml Shistosoma salt solution, and then diluted with salt solution to a concentration of 0.1 mg/lOO ml; for use, 1 ml is added to 11 medium. **Yeast extract/serum: yeast extract is made by combining 4 g Fleishmann's dry yeast with 16 ml Schistosoma salt solution and 16 ml FBS; the suspension is homogenized with a Sorvall Omni-Mixer (Sorvall) for 3 min, and then centrifuged at 1,000 g for 10 min; the supernatant is removed, and sterilized by filtration. ***Cholesterollalbumin: a suspension of cholesterol is prepared by adding 0.015 and 0.6 g bovine albumin to 60 ml of a selected reference medium; this is incubated for 1 h at 56°C with frequent agitation. The suspension is sterilized by filtration, which removes all but the submicroscopic particulate material; three drops of the filtrate are added to each culture at the time of preparation and again at each change of medium.

Medium 199 (= TC-199 medium) with Hank's salts (Morgan et al., 1950) The concentrations are given as mg/l,OOO ml culture medium. NaCl 8,000, KCl 400, MgS04·7H20 200, Na2HP04·2H20 60, KH2P04 60, CaCl2 140, NaHC03 350, glucose 1,000, phenol red 20, L-arginine hydrochloride 70, L-histidine hydrochloride 20, L-lysine monohydrochloride 70, DL-tryptophan 20, DL-phenylalanine 50, DL-serine 50, DL-methionine 30, DL-threonine 60, DL-leucine 120, DL-isoleucine 40, DL-valine 50, DL-glutamic acid monohydrate 50, DL-aspartic acid 60, DL-a-alanine 50, Lproline 40, L-hydroxyproline 10, glycine 50, L-glutamine 100, L-cystine 20, L-tyrosine 40, L-cysteine hydrochloride 0.1, sodium acetate 50, adenine 10, guanine hydrochloride 0.3, xanthine 0.3, hypoxanthine 0.3, uracil 0.3, thymine 0.3, sodium a-tocopherol phosphate 0.Q1, thiamin hydrochloride 0.Q1, pyridoxine hydrochloride 0.025, riboflavin 0.01, pyridoxal hydrochloride 0.025, niacin 0.025, niacinamide 0.025, calcium pantothenate 0.01, i-inositol 0.05, ascorbic acid 0.05, folic acid O.OI,p-aminobenzoic acid 0.05, biotin 0.01, menadione 0.01, glutathione 0.05, vitamin A 0.1, calciferol 0.1, cholesterol 0.2, Tween-80 20, adenosine triphosphate sodium salt 10, adenylic acid 0.2, D-2-desoxyribose 0.5, D-ribose 0.5, choline chloride 0.5, and ferric nitrate 0,1.

Medium 199 modified by Heacox et al. (Heacox et al., 1983) The concentrations are given as mg/l,OOO ml culture medium. NaCI17,550, CaCI2·2H20 930, KC1510, MgS04· 7H20 10,240, NaHC03 340, glucose 1,000, galactose 500, fucose(D) 250, fucose(L) 25, essential amino acids, non-essential amino acids and vitamin of Medium 199 as indicated in the instructions for the preparation of Medium 199, glutamine 100, gentamicin 1.0, penicillin 300,000 IU, Nereis coelomic fluid ultrafiltrate 0-20 ml, and Nereis vitellin 0 or 500.

Medium C for snail cell culture (Basch and Diconza, 1973) The concentrations are given as mg/l,OOO ml culture medium. NaCll,300, KCl 97, MgCI2·6H20 325, Na2HP04 17, CaCI2·2H20 558, NaHC03 1,769, Llysine 5.8, L-histidine 2.0, L-arginine 2.4, L-aspartic acid 5.6, L-threonine 6.2, L-serine 12.0, L-glutamic acid 9.4, L-proline 5.8, glycine 4.8, L-alanine 4.8, L-valine 3.2, L-methionine 0.03, L-isoleucine 2.0, L-Ieucine 2.9, L-phenylalanine 1.8, diphosphopyridine nucleotide 7.0, triphosphopyridine nucleotide 2.0, coenzyme A 2.5, cocarboxylase 2.0, flavin-adenine dinucleotide 2.0, biotin 1.0, folic acid 1.0, choline chloride 1.0, nicotinamide 1.0, D-calcium pantothenate 1.0, pyridoxal hydrochloride 1.0, thiamine hydro-


chloride 1.0, riboflavin 0.1, inositol 1.8, vitamin B12 10.0, nicotinic acid 2.0, p-aminobenzoic acid 2.0, pyridoxine hydrochloride 2.0, uridine 2.0, deoxyadenosine 10.0, deoxycytidine 10.0, deoxyguanosine 10, thymidine 10.0, cytosine 1.0, glucuronolactone 1.8, glutathione 10.0, phenol red 20.0, and glucose 100.0, made up to 1,000 ml with distilled water. The pH of the medium should be adjusted to 7.2-7.4 with 1 N KOH and 1 N HCl.

Medium X for Drosophila imaginal discs (Davis and Shearn, 1977) The concentrations are given as mg/1,000 ml culture medium. MgS04·7H20 1,230, MgCI2·6H20 3,192, KH2P04 200, K2HP04 256, KC1446, KHC03 300, potassium citrate 312, potassium fumarate 800, potassium a-ketoglutarate 870, potassium acetate 750, sodium citrate 3,122, sodium 1,4-piperazine diethanesulfonate (PIPES) 3,604, calcium succinate monohydrate 518, lactalbumin hydrolysate 9,000, L-proline 1,000, L-tryptophan 100, ~-alanine 50, L-cysteine 100, L-serine 250, y-aminobutyric acid 70, Bacto-peptone 5,000, trehalose 4,000, glucose 4,400, L-a-phosphatidylcholine, dimyristoyl1.0, L-a-phosphatidylcholine, distearoyl1.0, cholesterol 2.6, TC-Yeastolate 2,000, vitamin A (trans-retinol) 0.1, vitamin E (o-a-tocopherol) 0.1, sodium a-glycerophosphate 1,620, calcium phosphorylcholine 1,290, phosphorylethanolamine 70, BSA fraction V 10,000, polyvinylpyrrolidone (MW = 10,000) 250, polyvinylpyrrolidone (MW = 40,000) 1,000, and polyvinylpyrrolidone (MW = 360,000) 9,000. The pH of the medium should be adjusted to 6.8 with 1 N KOH and 1 N HCl. This medium can be used following supplementation with Altosid (a juvenile hormone analog; Zoecon) at a concentration of 31.0 ng/lOO ml medium and bovine insulin at 0.04 units/ 1 00 ml medium. Furthermore, 100 ml of this medium is conditioned either by preincubation with the fat bodies of five larvae or by co culturing of the discs with fat bodies.

MGM-434, MGM-443, MGM-448, MGM-450 and MGM-464 MGM-434 (chemically defined medium) The concentrations are given as mg/1,OOO ml medium. NaH2P04·2H20 958, NaHC03 292,KCl1,875,CaCI2 833, MgCI2·6H20 1,917, MgS04·7H20 2,333, glucose 3,333, fructose 417, sucrose 22,080, malic acid, 558, a-ketoglutaric acid 308, succinic acid 50, fumaric acid 46, L-a-alanine 263, ~-alanine 167, L-arginine hydrochloride 583, L-asparagine 293, L-aspartic acid 293, L-cystine 2l, L-glutamic acid 500, L-glutamine 500, glycine 542, L-histidine 2,083, L-isoleucine 42, L-Ieucine 63, Llysine hydrochloride 521, L-methionine 42, L-phenylalanine 125, L-proline 292, oL-serine 917, L-threonine 146, L-tryptophan 83, L-tyrosine 42, L-valine 83, thiamine hydrochloride 0.16, riboflavin 0.16, pyridoxine hydrochloride 0.16, niacin 0.16, calcium pantothenate 0.16, biotin 0.08, folic acid 0.16, isoinositol 0.16,p-aminobenzoic acid 0.16, choline chloride 1.6, polyvinylpyrrolidone K-90 500. The pH of the medium should be adjusted to 6.5 with concentrated KOH. MGM-443 (Mitsuhashi, 1980) This is MGM-434 medium modified by the addition of 10% FBS. The pH of the medium is adjusted to 6.3 with concentrated KOH solution. MGM-448 (Mitsuhashi, 1984) This is MGM-443 medium fortified by the addition of inosine 200 mg/l, cytochrome c 100 mg/I, fetuin 20 mg/l, and bovine plasma albumin fraction V 10,000 mg/l, FBS 100 mll

414 Appendix

1, or Antheraea pernyi hemolymph 30 mlll. The pH of the medium is adjusted to 6.3 with concentrated KOH solution. MGM-450 (Mitsuhashi and Inoue, 1988) This medium is prepared by adding 3,000 mg tryptose phosphate broth to 1,000 ml of MGM-448 medium. The pH is adjusted to 6.3 with concentrated KOH solution. This medium can also be used by adding FBS at 10%, or FBS 5% and Antheraea pernyi hemolymph 3%. MGM-464 (Mitsuhashi, 2001) This medium is prepared by adding 3,000 mg tryptose phosphate broth, 3,000 mg TCYeastolate (Difco), and 3,000 mg lactalbumin hydrolysate to 1,000 ml of MGM-448 medium. The pH of the medium is adjusted to 6.3 with concentrated KOH solution. This medium is used by adding FBS at 20%.

MM medium (Mitsuhashi and Maramorosch, 1964) The concentrations are given as mg/l,OOO ml culture medium. NaCl 7,000, KCl 200, CaC12·2H20 200, MgC12·6H20 100, NaH2P04 200, NaHC03 120, glucose 4,000, lactalbumin hydrolysate 6,500, TC-Yeastolate 5,000, and FBS 0-200 ml, made up to 1,000 ml with distilled water. The pH is adjusted to 6.3 with KOH (the pH of the medium is already around 6.3 without any adjustment). The medium can be sterilized before the addition of FBS by autoclaving at 121°C for 15 min. The medium can be used as a serum-free medium (= MM-SF medium).

Nudibranch culture medium (Tamse et aI., 1995) This is Leibovitz's L-15 medium supplemented with 444 mM NaCl , 10.0 mM KCl, 28.0 mM MgC12·6H20, 27.0 mM MgS04·7H20, 2.0 mM NaHC03, 35.0 mM D-glucose, 15.0 mM HEPES, 11.0 mM CaC12·2H20, penicillin 50 lUI ml, and streptomycin 50 Ilg1 ml. The pH of the medium should be 7.4-7.5.

Ogura's medium (Ogura, 1989) The concentrations are given as mg/l,OOO ml culture medium. NaCl 7,900, KCl 290, CaClz·2H20 200, MgS04·7H20 150, Na2HP04·12Hp 380, KH2P04

80, NaHC03 1,200, glucose 1,500, lactalbumin hydrolysate 500, TC-Yeastolate 250, chicken embryo extract 2.5 ml, nematode hemolymph * 5 ml, penicillin 105 IV, and streptomycin 100, made up to 1,000 ml with distilled water. *Nematode hemolymph is prepared from the female adults of Toxocara canis.

Oncomelania culture medium (Iwanaga et aI., 1985) Vnless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium. NaC12,800, KCl150, Na2 HP04 70, MgS04·7H20 450, CaC12·2H20 530, NaHC03 50, galactose 5,000, glucose 5,000, trehalose 1,000, a-ketoglutaric acid 1,200, fumaric acid 800, malic acid 800, succinic acid 800, L-lysine hydrochloride 208, L-histidine 100, Larginine hydrochloride 144, L-aspartic acid 532, L-threonine 288, L-serine 360, L-glutamic acid 516, L-proline 248, glycine 388, L-alanine 352, L-valine 104, L-methionine 88, Lisoleucine 116, L-leucine 292, L-tyrosine 128, L-phenylalanine 156, Eagle's MBE vitamin (xl00; Gibco) 40 ml, and phenol red 5, made up to 1,000 ml with distilled water. Vse the medium by adding heat-treated horse serum at a concentration of 10%. The pH of the medium should be 7.0-7.2.


Pf-35 medium (Machii and Wada, 1989) Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium. NaCI26,220.0, KCll,080.0, MgS04 3,130.0, MgCl2 2,200.0, CaCl2 1,120.0, NaHC03 300.0, NaH2P04 44.0, L-cystine 0.7, L-tyrosine 3.6, taurine 128.9, L-aspartic acid 12.0, L-threonine 3.6, L-serine 9.5, L-asparagine 5.3, L-glutamic acid 19.1, L-glutamine 17.5, L-proline 3.5, glycine 15.0, L-a-alanine 17.8, a-amino-n-butyric acid 0.8, L-valine 4.7, L-methionine 3.0, L-isoleucine 2.6, L-Ieucine 3.9, L-phenylalanine 1.7, L-ornithine 0.9, L-Iysine 7.3, L-tryptophan 1.0, L-histidine 4.7, L-arginine 17.4, phosphoserine 1.0,p-aminobenzoic acid 0.05, biotin 0.01, calcium pantothenate 1.0, choline chloride 250.0, folic acid 0.01, inositol 0.05, niacin 0.03, niacinamide 5.0, pyridoxal hydrochloride 0.025, pyridoxine hydrochloride 1.0, riboflavin 1.0, cyanocobalamin 0.01, thiamine hydrochloride 10.0, ascorbic acid 40.0, vitamin A 0.1, a-tocopherol acetate 0.01, calciferol 0.1, menadione 0.01, adenine 10.0, guanine hydrochloride 0.3, hypoxanthine 0.3, thymine 0.3, uracil 0.3, xanthine 0.3, adenylic acid 0.2, 2-deoxy-o-ribose 0.5, o-ribose 0.5, Tween-80 5.0, cholesterol 0.2, glutathione 0.05, sodium acetate 50.0, glucose 300, ATP 10.0, lactalbumin hydrolysate 100.0, phenol red 5.0, penicillin 200, kanamycin 100, and FBS 100 ml, made up to 1,000 ml with distilled water.

Planarian cell culture medium (Betchaku, 1967) Unless otherwise indicated, the concentrations are given as mgll,OOO ml culture medium. L-Arginine hydrochloride 5.6, L-cystine 2.0, L-histidine 1.1, L-isoleucine 8.7, L-Ieucine 4.7, L-Iysine 6.1, L-methionine 2.7, L-phenylalanine 2.8, L-threonine 4.0, L-tryptophan 0.7, L-tyrosine 6.0, L-valine 3.9, L-glutamine 48.9, choline chloride 0.3, nicotinamide 0.3, calcium-o-pantothenate 0.3, pyridoxal hydrochloride 0.3, riboflavin 0.03, thiamine hydrochloride 0.3, i-inositol 0.3, biotin 0.3, folic acid 0.3, sodium pyruvate 150.0, glucose 300.0, calf serum 3 ml, NaCll,750.0, KCI25.0, CaCl2 50.0, NaHC03 100.0, neomycin 100.0. The pH of the medium should be adjusted to 7.2-7.4 with 1 N KOH and 1 N HCl.

Rasmont medium M (Rasmont, 1961) The concentrations are given as milliequivalent II. Ca++ 2.00, Mg++ 1.00, Na+ 1.00, K+ 0.05, Cl~ 2.05, S04 1.00, Si03~~ 0.50, HC03~ 0.50.

RML-375 medium (Moulton, 1978) AI: 1 : 2 mixture of Eagle's MEM, Hank's BSS and Leibovitz's L-15 medium is supplemented with 20% FBS (heat-inactivated for 1 h at 56°C), 0.1% bovine albumin, 10% tryptose phosphate broth stock solution, made by dissolving 109 tryptose phosphate broth in 100 ml distilled water, and 50 glml gentamicin.

S-20 medium (Landureau and Grellet, 1972) The concentrations are given as mg/1,OOO ml culture medium. NaCl 8,474, KCI 7,070, MgS04·7H20 1,016, CaCl2 222, MnS04·H20 32, P03H3 900, glucose 4,000, L-arginine hydrochloride 800, L-aspartic acid 506, L-cysteine hydrochloride 646, L-glutamic acid 1,500, glycine 750, L-histidine 202, L-isoleucine 131, L-Ieucine 197, L-Iysine hydrochloride 183, L-methionine 254, L-phenylalanine 99, L-proline 750, L-serine 32, L-threonine 100, L-tryptophan 102, L-tyrosine 90, L-valine 94, folic acid 0.02, d-biotin 0.01, choline chloride 181.48, cyanocobalamine 0.04, inositol 0.05, nicotinamide 0.04, pyridoxine hydrochloride 0.04, riboflavin 0.19, thiamine hydrochloride 0.67, cal-

416 Appendix

cium pantothenate 0.24, made up to 1,000 ml with distilled water. The pH of the medium should be adjusted to 6.7 with 1 N KOH and 1 N HCl.

Sengel's medium (Sengel, 1960) Salt solution (Holtfreter's solution, 1931): NaC13,500 mg, KCI SO mg, CaCl22H20 100 mg, and NaHC03 200 mg, made up to 1,000 ml with distilled water. Culture medium: 5 parts of 1 % agar in the above salt solution, 4 parts of 0.1 % glucose in the above salt solution, and 3 parts chicken embryo (9 days old) extract diluted to 50% with the above salt solution.

SH-7 medium (Furuta and Shimozawa, 1983) Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium. NaCI 2,483.0, KCI 496.0, CaCl2 6.0, MgCl2 152.0, NaHC03 1,769.0, glucose 50.0, trehalose 50.0, L-a-alanine 41.2, L-arginine hydrochloride 6.1, L-asparagine monohydrate 14.5, L-aspartic acid 3.1, L-cystine 3.3, L-glutamic acid 3.7, L-glutamine 3.7, glycine 3.0, L-histidine dihydrochloride 4.1, L-isoleucine 3.3, L-Ieucine 2.3, L-Iysine hydrochloride 5.8, L-phenylalanine 5.7, L-serine 11.8, L-threonine 27.2, L-tyrosine 2.8, L-valine 4.9, thymidine 0.35, uridine 22.4, hypoxanthine 2.0, tricine 2,750.0, putrescine 0.15, phenol red 2.0, D-biotin 0.007, choline chloride 7.0, calcium pantothenate 0.25, folic acid 0.5, inositol 9 .0, pyridoxal hydrochloride 0.03, riboflavin 0.02, thiamine hydrochloride 0.15, cyanocobalamin 0.5, nicotinamide 0.02, linoleic acid 0.04, a-lipoic acid 0.1, FBS (dialyzed and inactivated) 100.0 ml, and chicken embryo extract 40.0 ml, made up to 1,000 ml with distilled water.

Shields and Sang's Medium (Shields and Sang, 1970) The concentrations are given as mg/l,OOO ml culture medium. MgS04·7H20 5,130, CaCI2·6H20 1,740, KCI3,130, NaC1860, NaH2P04·2H20 880, KHC03

180, monosodium malate·2H20 950, monosodium a-ketoglutarate 420, disodium fumarate 80, disodium succinate·6H20 140, monosodium L-glutamate 2,460, L-aspartic acid ISO, L-threonine 500, L-serine 350, L-asparagine 300, L-glutamine 600, L-proline 400, glycine 500, L-a-alanine 1,650, L-valine 420, L-methionine 120, L-isoleucine 270, Lleucine 400, L-tyrosine 260, L-phenylalanine 240, ~-alanine 100, L-histidine 550, L-tryptophan 100, L-arginine 500, L-Iysine 680, L-cystine 200, L-cysteine 800, glutathione 5, glucose 4,600, and TC-Yeastolate (Difco Laboratories) 2,000. The pH of the medium is adjusted to 6.9 with 1 % NaOH. Use the medium by adding 10% FBS.

Shields and Sang's medium modified by Poels (1972) The concentrations are given as mg/l,OOO ml culture medium. NaCI 860, NaH2P04·2H20 880, KCI 3,130, KHC03 180, CaCl2·2H20 1,160, MgS04·7H20 5,130, monosodium malate·2H20 950, monosodium a-ketoglutarate 420, disodium succinate·6H20 140, disodium fumarate 80, monosodium pyruvate 20, monosodium Lglutamate 760, L-aspartic acid 90, L-threonine 150, L-serine 175, L-asparagine 150, Lglutamine 300, L-proline 400, glycine 175, L-a-alanine 750, L-valine 90, L-methionine 150, L-isoleucine SO, L-Ieucine 90, L-tyrosine 30, L-phenylalanine 30, ~-alanine 120, Lhistidine 190, L-tryptophan 100, L-arginine 210, L-Iysine 70, L-cystine 10, L-cysteine 60, TC-Yeastolate 2,000, glucose 300, trehalose 800, and FBS 100 ml. The pH of the medium is adjusted to 6.9 with 1 N NaOH.


Shields and Sang's medium modified by Sinha and Lakhotia (1980) Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium. NaCl 860, NaHzP04·2HzO 8S0, KCl 3,130, KHC03 ISO, CaClz·2HzO 1,160, MgS04·7HzO 5,130, malic acid 950, a-ketoglutaric acid 420, fumaric acid SO, succinic acid 140, pyruvic acid 20 Ill, glucose 300, trehalose SOO, lactalbumin hydrolysate S,120, Bacto-peptone 5,000, yeast extract paste 2,000, streptopenicillin 1,000, insulin 0.4 units. The pH of the medium is adjusted to 7.0 with I N KOH and 1 N HCl. For maintaining the salivary glands of Drosophila, preconditioning of the medium or co culture with fat bodies provides better results.

Snail organ culture medium (Wattez, 1975) Stock solution 1: Agar 1% (Difco Laboratories) in Gey's solution mixed with the same amount of peptone (Difco Laboratories) Stock solution 2: mix 63 parts TC-199, S5 parts of 0.3% sodium bicarbonate, S50 parts Hank's solution, and 2 parts yeast extract. Mix 7 drops of stock solution 1 and 6 drops of stock solution 2, supplement this mixture with penicillin G 100 IU and polymyxin B 25 IU. The pH of the medium is adjusted to 7.45 with sodium bicarbonate.

TC-199 (see Medium 199).

TC-199MK (Mcintosh et al., 1973) This medium consists of a mixture of equal parts of TC-199 with Hanks' salts and Melnick's monkey kidney medium "N', both without sodium bicarbonate. To this, 10% inactivated FBS is added. Antibiotics are incorporated at final concentrations of 50 IU / ml penicillin and 50 Ilg/ml streptomycin. 1 ml/lOO ml medium of 200 mM glutamine is added to the final product. The pH of the medium is 6.9 without adjustment. The osmolarity is 266 mOsmollkg.

TTP medium (Azumaya and Teshirogi's medium) for planarians (Teshirogi, 1998) Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium. NaCll,S59.S, KCl 26.4, CaClz 53.6, MgS04·7HzO 90.1, MnClz·4HzO 0.3, NaHC03 SOO.O, L-aspartic acid I.S, L-threonine 6.2, L-serine 6.0, L-glutamic acid 7.7, L-glutamine 4S.9, glycine 1.3, L-alanine 5.3, L-cystine 2.0, L-valine 3.1, L-methionine 2.0, L-isoleucine 1.5, L-leucine I.S, L-tyrosine 1.2, L-phenylalanine 2.S, L-lysine 7.7, L-histidine 4.1, L-tryptophan 0.7, choline chloride 0.3, nicotinamide 0.3, calcium-o-pantothenate 0.3, pyridoxal hydrochloride 0.3, riboflavin 0.03, thiamine hydrochloride 0.3, i-inositol 0.3, biotin 0.3, folic acid 0.3, glucose 300.0, trehalose 50.0, tricine 2.5, phenol red 2.0, and FBS (heated at 56°C for 30 min) 10.0 ml. The pH of the medium is adjusted to 7.4-7.6 with 1 N KOH and 1 N HCl. The osmolarity of the medium is approximately SO mOsmollkg.

Wolff et al?s medium C (Wolff et al., 1953) First, prepare solution C by dissolving following the chemicals in 25 ml Tyrode's solution (see "Salt Solutions" in Appendix I): L-lysine hydrochloride 15.1 mg, L-arginine 7.7 mg, DL-methionine 2.6 mg, oL-histidine hydrochloride 3.1 mg, L-glutamic acid 14.1 mg, DL-aspartic acid 5.9 mg, L-proline 5.1 mg, L-cysteine hydrochloride 1.5 mg, Ltryptophan 5 mg, OL-phenylalanine 7 mg, L-leucine 9 mg, OL-isoleucine 10 mg, OL-threo-

418 Appendix

nine 12 mg, DL-valine 14 mg, glycine 2.5 mg, L-tyrosine 1 mg, L-hydroxyproline 2.5 mg, DL-serine 12.5 mg, taurine 5 mg, ornithine dihydrochloride 5 mg, and L-asparagine 5 mg. Adjust the pH to 7.5 with 1 N KOH and 1 N HC!. To prepare medium C, dissolve 0.25 mg glutathione into solution C.

Wyatt's medium (Wyatt, 1956) The concentrations are given as mg/l,OOO ml culture medium. NaH2P04 1,100, KCl 2,980, MgCl2·6H20 3,040, MgS04·7H20 3,700, CaCl2 810, glucose 700, fructose 400, sucrose 400, malic acid 670, a-ketoglutaric acid 370, succinic acid 60, fumaric acid 55, DL-a-alanine 450, ~-alanine 200, L-arginine 700, L-asparagine 350, Laspartic acid 350, cysteine 80, L-cystine 25, L-glutamic acid 600, L-glutamine 600, glycine 650, L-histidine 2,500, DL-isoleucine 100, DL-leucine 150, DL-lysine 1,250, DL-methionine 100, L-phenylalanine 150, L-proline 350, DL-serine 1,100, DL-threonine 350, Ltryptophan 100, L-tyrosine 50, and DL-valine 200. The pH of the medium is adjusted to 6.35 with 1 N KOH and 1 N HC!. Use the medium by adding heat-treated Bombyx mori hemolymlph.

ZH 1% medium (Wyss, 1977) Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium. CaCl2·2H20 890, KCl 2,000, MgCl2·6H20 1,000, MgS04·7H20 3,700, NaCl 2,050, NaH2P04·2H20 470, CuS04·5HzO 0.01, FeS04·7Hp 0.6, MnS04·7H20 0.3, ZnS04·7H20 0.6, L-arginine 170, L-asparagine l30, L-cysteine hydrochloride 310, L-glutamine 700, Lglutamic acid 290, glycine 750, L-histidine 150, L-isoleucine l30, L-leucine l30, L-lysine 180, L-methionine 150, L-phenylalanine 160, L-proline llO, L-serine 520, L-threonine 120, L-tryptophan 100, L-tyrosine 90, L-valine 120, glucose 2,000, malic acid 670, oxaloacetic acid 250, sodium acetate·3H20 25, succinic acid 60, biotin 0.01, calcium pantothenate 0.02, choline chloride 0.2, folic acid 0.02, m-inositol 0.02, niacin 0.02, niacinamide 0.02, pyridoxine hydrochloride 0.02, pyridoxal hydrochloride 0.02, riboflavin 0.02, thiamine hydrochloride 0.02,p-aminobenzoic acid 0.02, cyanocobalamin 0.1, phenol red 10, high molecular weight fraction of yeast extract* 1,500 (equivalent to original yeast extract), and heat-inactivated horse serum 10 m!. The pH is adjusted to 6.6 with NaOH. *The fractionation of yeast extract was made using Sephadex G25 (Sigma) chromatography, and fractions 23-46 are used (Wyss, 1977).

ZW medium (Wyss, 1982) Unless otherwise indicated, the concentrations are given as mg/1,000 ml culture medium. L-a-Alanine 45, ~-alanine 45, L-arginine 174, L-asparagine 150, L-aspartic acid l33, Lcysteine hydrochloride 315, L-glutamine 680, L-glutamic acid 294, glycine 750, L-histidine 620, L-hydroxyproline 65, L-isoleucine l31, L-leucine l31, L-lysine hydrochloride 182, L-methionine 149, L-ornithine hydrochloride 84, L-phenylalanine 165, L-proline 115, L-serine 525, L-threonine 119, L-tryptophan 102, L-tyrosine 90, L-valine 117, glucose 2,000, inosine 2.7, thymidine 2.4, uridine 2.4, 1-thioglycerol 5.4, DL-carnitine hydrochloride 1,000, reduced glutathione 50, malic acid 670, oxaloacetic acid 250, sodium acetate·3Hp 25, sodium pyruvate llO, succinic acid 60, CaCl2·2H20 147, KCl


2,000, MgS04·7H20 1,230, NaCI3,200, NaH2P04·H20 420, CuS04 0.001, FeS04·7H20 1.0, MnS04·7H20 1.0, ZnS04·7H20 1.0, biotin 0.05, calcium pantothenate 5.0, choline chloride 50.0, folic acid 0.05, m-inositoI50.0, niacinamide 0.5, pyridoxal hydrochloride 5.0, riboflavin 0.01, thiamine hydrochloride 5.0, cyanocobalamin 0.05, cholesterol 0.01, linoleic acid 0.01, and phenol red 10.0, made up to 1,000 ml with distilled water. The pH of the medium is adjusted to 6.6 with 1 N NaOH. The final osmolarity of the medium is 250 mOsmollkg.

References

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Appendix 2

List of Reported Continuous Invertebrate Cell Lines

How many invertebrate cell lines exist is uncertain, because there are many cell lines that have not been reported properly, and some of the reported cell lines have been lost already. There are also some cell lines that are known to be mislabeled or cross-con-taminated. The following list contains only properly reported cell lines; cell lines that no longer exist or those that have been ascertained as mislabeled are marked with asterisks.

Species Stage Tissue Designation Report

Arthropoda Insecta Blattaria Phyllodromiidae Blabera fusca 'Embryo Whole Landureau, 1968

Blattella germanica Embryo Whole UM-BGE-1, -2, Kurtti and Brooks, -4, and-5 1977

B. germanica Embryo Whole Landureau, 1966 Blattidae Gromphadorhina Larva Dorsal Quiot et ai., 1985

laevigata vessel

Periplaneta Embryo Whole EPa Landureau, 1968 americana

p. americana Larva Hemocyte HPa-33 Landureau and -34 and Grellet, 1975

P. americana Adult Fat body APa Philippe, 1982 Coleoptera Cerambycidae Xylotrechus Larva Fat body XP-1 Iwabuchi, 1999 pyrrhoderus Chrysomelidae Diabrotica Embryo Whole IPLB-DU182A, Lynn and

undecimpunctata IPLB-DU182E Stoppleworth, 1984

Leptinotarsa Embryo Whole ZIZ-LD-1 Dubendorfer and decemlineata Liebig, 1992

L. decemlineata Embryo Whole IPLB-CPB2 Lynn, 1995 Culcurionidae Anthonomus grandis Embryo Whole AGE,AGF Barcenas et ai., 1989 A. grandis Embryo Whole BRL-AG-1,-2, Stiles et ai., 1992

-3A, -3C, and -4

422 Appendix


Scarabaeidae

Antitragus parvulus Embryo Whole APE-1 Fernon et aI., 1996

A. parvulus Larva Hemocyte APHL-1 Fernon et aI., 1996

Anomala costata* Embryo Whole FRI-AnCo-5B Mitsuhashi, 1989

Heteronychus arator Embryo Whole DSIR-HA-1l79 Crawford, 1982 Diptera Ceratopogonidae Cu lico ides Embryo Whole CuVa-1-4 Wechsler et al.,1989

variipennis Culicidae Aedes aegypti* Matured larva Whole Grace, 1966

A. aegypti* Neonate larva Whole ATC-lO Singh, 1967

A. aegypti Embryo Whole 59,364 Peleg,1969

A. aegypti Neonate larva Whole Mos.20, 20A, and 29 Varma and Pudney, 1969

A. aegypti Larva Whole Mill Hill Bailey et aI., 1975

Aedes albopictus Neonate larva Whole ATC-15 Singh, 1967

A. albopictus Neonate larva Whole ATC-89 and -91 Bhat and Guru, 1973

A. albopictus Neonate larva Whole Hsu et aI., 1978

A. albopictus Neonate larva Whole NIAS-AeAI-2 Mitsuhashi, 1981a

Aedes dorsalis Neonate larva Whole Cahoon et aI. 1978

Aedes krombeini Neonate larva Whole NIVI-AK-453 Pant et aI., 1992

Aedes malayensis Neonate larva Whole Mos60 Varma et aI., 1974

Aedes novalbopictus Neonate larva Whole ATC-173 and -176 Bhat and Guru, 1973

Aedes Neonate larva Whole Mos 61 Varma et aI., 1974 pseudoscutellaris

Aedes triseriatus Neonate larva Whole Rowley et aI., 1984

Aedes vexans * Neonate larva Whole Sweet and McHale, 1970

Aedes vittatus Neonate larva Whole ATC-121 Bhat and Singh, 1970

Aedes w-albus Neonate larva Whole ATC-136 and -137 Singh and Bhat, 1971

Anopheles gambiae Neonate larva Whole Mos 55 Marhoul and Pudney,1972

Anopheles stephensi Neonate larva Whole Schneider, 1969

A. stephensi Neonate larva Whole Mos43 Pudneyand Varma, 1971

A. stephensi Neonate larva Whole Mos 44, 45, 46 Pudney et aI., 1973 Cliseta inornata* Neonate larva Whole Sweet and McHale,

1970

Culex Embryo Whole ATC-415 Pant and Dhanda, bitaeniorhynchus 1980

Culex molestus Adult Ovary CmOv Kitamura, 1970

List of Reported Continuous Invertebrate Cell Lines 423


Culex pipiens Embryo Whole cp-73 Chao and Ball, 1973

Culex quinquefasciatus Adult Ovary Hsu et aI., 1970

Culex salinarius Neonate larva Whole Schneider, 1973

Culex Neonate larva Whole Schneider, 1973 tritaeniorhynchus

C. tritaeniorhynchus Adult Ovary Hsu et aI., 1972 summorosus

Haemagogus equinus Neonate larva Whole GML-HE-12 Oro, 1984

Toxorhynchites Neonate larva Whole 7A-42 Tesh,1980 amboinensis

T. amboinensis Embryo Whole TA-9 Tesh,1980

T. amboinensis Larva Whole Tra-1 to -22 Kuno,1980

T. amboinensis Neonate larva Whole Tra-147, -272, Kuno, 1981 and -293

T. amboinensis Larva Whole RU-Tae-12V Munderloh et al., 1982 Chironomidae

Camptochironomus Embryo Whole Wyss, 1982 tentans

Drosophilidae Drosophila hydei Embryo Whole KUN-DH-14,-15, Sondermeijer et aI.,

-33, and -47 1980

Drosophila Neonate Whole WR-72-Di-1 Schneider and immigrans Larva Blumenthal, 1978

Drosophila Embryo Whole Kc Echalier and melanogaster Ohanessian, 1969

D. melanogaster Embryo Whole 67j25 Kakpakov et aI., 1969

D. melanogaster Embryo Whole 68C and 68K Echalier and Ohanessian, 1970

D. melanogaster Embryo Whole S1, S2, and S3 Schneider, 1972

D. melanogaster Embryo Whole GM1,GM2, Mosna and Dolfini, and GM3 1972

D. melanogaster Embryo Whole 3XIII, 1.59, Mosna and and 0.57 Barigozzi, 1972

D. melanogaster Embryo Whole IHI Paradi, 1972

D. melanogaster Embryo Whole B,G,I,andJ Richard-Molani, 1975

D. melanogaster Embryo Whole 0.57 and 1.59 Mosna and Barigozzi, 1976

D. melanogaster Embryo Whole 75e7vgl,75e7vg2, Kakpakov et aI., 75e7vg3,75e7vg4, 1977 75e7vg6, and 75e7vg7

D. melanogaster Embryo Whole ORE-R, B26ywf, Petersen et aI., 1977 and Igl4

424 Appendix


D. melanogaster Embryo Whole 1182-1 to -7 Debec,1978

D. melanogaster Embryo Whole Er1(a,b,c,d1, Regenass and d2,e) and Er36 Bernhard, 1979

D. melanogaster Embryo Whole EOR,ECS, Bernhard et al., EFM1 Eml, EE, 1980 and Ealdox

D. melanogaster Embryo Whole 3P80, 5P80, p80, Halfer et aI., 1980 11P80AB,12P80D, SP102, 8P102, 2P102, lOP 102, 11P102, and lSP102

D. melanogaster Embryo Whole UCD-Dm-w1, Hirschi and Boyd, and 15P102 1981

D. melanogaster Embryo Whole 1.56,3.38, lXII, Mosna,1983 and 0.57

D. melanogaster Embryo Whole WT-CS3, -CS-5, Simcox et al.,1985 and WT-LM6

D. melanogaster Embryo Whole ML82-8 Todo et aI., 1985

D. melanogaster Embryo Whole SSl,SS13, Chalvet et aI., 1998 and SS14A

D. melanogaster Larva Tumorous 1(2)mbn and Gateff et aI., 1980 hemocyte 1(3)mbn

D. melanogaster Larva Imaginal disc ML-DmD1 to D20 Ui et aI., 1987

D. melanogaster Larva Imaginal CMEW1 to 2, Cullen and Milner, disc and CME L1 to 2 1991

D. melanogaster Larva Nervous ML-DmBG 1 to 8 Ui-Tei et aI., 1994 system

Drosophila simulans Embryo Whole ML83-8b Yoshioka et aI., 1992

Drosophila virilis Neonate larva Whole WR-75-Dv-1 Schneider and Blumenthal, 1978

D. virilis Embryo Whole 79f7Dv3g Braude Zolotarjova et aI., 1986

Muscidae

Musca domestica Embryo Whole Eide,1975

Psychodidae Lutzomyia Embryo Whole LL-5 Tesh and Modi,

longipalpis 1983

Sarcophagidae

Sa rcophaga Embryo Whole NIH-SaPe-4 Takahashi et al. peregrina 1980

Tipulidae

Tipula paludosa Larva Hemocyte Griffiths and Carter, 1982



Trypetidae Ceratitis capitata Embryo Whole CEC-CC-128 Cavalloro, 1981

and -130 Heteroptera Cimicidae

Cimex hemipterous Embryo Whole NIVI-CH-440, Pant et al., 1988 -442, and -445

Homoptera Aleyrodidae

Bemisia tabaci Embryo Whole Btb(Ba)97Hunter- Hunter and Polston, Polston 2001

Jassidae

Aceratagallia Embryo Whole AS-1 Chiu and Black, 1969 sanguinolenta

Agallia constricta Embryo Whole AC20 Chiu and Black, 1967

Agallia Embryo Whole Chiu and Black, 1967 quadripunctata

Colladonus Richardson and montanus Jensen, 1971

Macrosteles Embryo Whole Peters and sexnotatus Spaansen, 1973

Nephotettix Embryo Whole NC-15, -19, -20, Kimura, 1984 cincticeps 21, -24, and -25

Nephotettix Embryo Whole NN-1, -2, -5, and-7 Kimura, 1985 nigropictus

Hymenoptera Aphidiidae Ephedrus plagiator Larva Whole IBT-EP-1 and-2 Eppler et ai., 1980

Diprionidae Neodiprion lecontei Embryo Whole FPMI -NL-2, -4, Sohi and Ennis,

-10, -18, -21, -22, 1981 -28, and -32

Trichogrammatidae Trichogramma Larva Whole IPLB-TpE-1 Lynn and Hung, 1986

pretiosum

Trichogramma Larva Whole IPLB-Tcon-l Lynn and Hung, 1991 confusum

Trichogramma Larva Whole IPLB-Tex-l Lynn and Hung, 1991 exiguum

Lepidoptera Arctiidae

Estigmene acrea Larva Hemocyte EA1174A and-H Granados and Naughton, 1975

Spilarctia Larva Fat body NIAS-SpSe-l Mitsuhashi, 1984 seriatopunctata

426 Appendix


Spilosoma imparilis Larva Fat body FRI-Splm -1229 Mitsuhashi and Inoue, 1988

Bombycidae

Bombyx mandarin a Embryo Whole SES-Bma-01 Ninaki et al., 1988

Bombyx mori* Larva Ovary Grace, 1967

B. mori Pupa Ovary S.P.C.Bm 36 Quiot,1982

B.mori Embryo Whole SES-BoMo -15A Inoue and Mitsuhashi, 1984

B.mori Embryo Whole SES-BoMo -J125 Imanishi and and -C129 Ohtsuki, 1988

B. mori Embryo Whole SES-Bm-e 21 and Ninaki et aI., 1988 SES-Bm-130

B. mori Embryo Whole TUAT-BME-ll Pandharipande, and -15 1994

B. mori Larva Midgut Baines et al., 1994

B.mori Embryo Whole NISES-BoMo -MK, Imanishi et aI., 1999 -KG, -DZ, and -OH

Gelechiidae Gnorimoschema Embryo Whole GOl-874 Pant et al. 1977

operculella

G. operculella Embryo Whole ORS-Pop-2, Lery et aI., 1997 -3, -4,and -93

Pectinophor Embryo Whole Lery et aI., 1996 gossypiella

Phthorimaea Embryo Whole ORS-Pop-93 Lery et aI., 1995 (= Gnorimoschema) operculella

Lasiocampidae Malacosoma disstria Larva Hemocyte IPRI108 Sohi, 1973a

Malacooma neustria Larva Hemocyte FRI-MntH Mi tsuhashi, 1996 testacea -52OA

Lymantriidae

Lymantria dispar Pupa Ovary SCLd-135 Quiot,1976

L. dispar Pupa Ovary IPLB-LD-64, Goodwin et al. -65, -66, and 67 1978

L. dispar Larva Midgut Baines et aI., 1994

Orgyia leucostigma Embryo Whole IPLB-OIE505 Lynn and Shapiro, 1997

Noctuidae

Anticarsia Embryo Whole UFL-AG-286 Sieburth and gemmatalis Maruniak, 1988

Euxoa scandens Pupa Ovary IAFEs-1 Belloncik et aI., 1985

Helicoverpa Adult Ovary CSIRO-BCIRL Mcintosh et al., 1999 armigera -HAl, 2,and 3



Helicoverpa Adult Ovary CSIRO-BCIRL McIntosh et al., 1999 punctigera -HPl, 2, 3, 4, and 5

Heliothis armigera Pupa Ovary ACIRL-HA McIntosh et aI., -AMI 1983

Heliothis virescens Pupa Ovary BCIRL-HV McIntosh et aI., -AM-l and 1981 AM-2

Heliothis zea Pupa Ovary Goodwin, 1975

H.zea Pupa Ovary BCIRL-Hz McIntosh and Ignoffo, -AMl,AM-2, 1981 and -AM-3

H. zea Pupa Ovary IPLB-Hz-II 0, Goodwin et aI., -1240, and -lO74 1982

H. zea Adult Ovary IMC-HZ-1 Hink and Ignoffo, 1970

Leucania separata Larva Fat body NIAS-LeSe-ll Mitsuhashi, 1983

Mythimna Larva Hemocyte SIE-MSH-805 Shen and Liu, 1983 (= Leucania) separata

Mamestra brassicae Larva + pupa Dorsal vessel Mb-1203 Miltenburger and + gonad David, 1976

M. brassicae Larva Hemocyte Mb-0503, -2006, Miltenburger and -2007, and -2506 David,1976

M. brassicae Pupa Ovary HP-B-MB Hilwig, 1976

M. brassicae Pupa Ovary NIAS-MB-19, Mitsuhashi, 1977 -25, and -32

M. brassicae Larva Hemocyte IZD-MB0503 Miltenburger et aI., -0504, -2006, 1977 -2007, and -1203

M. brassicae Larva Fat body NIAS-MaBr-85 Mitsuhashi,1981b

M. brassicae Larva Hemocyte NIAS-MaBr-92 Mitsuhashi and and -93 Shozawa, 1985

M. brassicae Larva Fat body SES-MaBr-1, -2, Inoue and -3, -4, and-5 Mitsuhashi, 1985

M. brassicae Neonate larva Whole MbL-3 Kondo et aI., 1995

Mythimna convecta Pupa Fat body BPMNU Khurad et aI., 1991 -MyCo-1

Pseudalletia Embryo Whole BTI-Pu-527 Wang et aI., 1999 unipuncta A7, -527B9,

-7202-527M, -527M1B, and 527A7S,

Spodoptera exigua Neonate larva Whole Se3FH, Se4FH, Hara et aI., 1993 Se5FH, Se6FHA, and Se6FHB

428 Appendix


S. exigua Neonate larva Whole UCR-SE-l Gelernter and Federici, 1986

Spodoptera frugiperda Pupa Ovary Goodwin et aI., 1975

S. frugiperda Pupa Ovary IPLB-SF-21 Vaughn et al., 1977 and -1254

S. frugiperda Larva Imaginal IAL-SHDI Lynn and disc Oberlander, 1983

Spodoptera littoralis Pupa Ovary UIV-SL-573 Knudson et aI., 1980

S. littoralis Pupa Ovary HPB-SL-26 Hilwig, 1980

Spodoptera litura Pupa Ovary TUAT-SpLi-221 Mitsuhashi, 1995

S. litura Larva Ovary NIV-SU-983 Pant et aI., 1997 and -992

Spodoptera Adult Ovary BCIRL-S03 Chen et al. 1984 ornithogalle -HNUI and

-S04-HNU2

Thysanoplusia Larva Fat Body TI-l Iwabuchi, 2000 intermixta

Trichoplusia ni Adult Ovary 8 TN-36 Hink,1970

T. ni Pupa Ovary Goodwin et aI., 1975

T. ni Larva Imaginal disc IAL-TNDI Lynn et aI., 1982

T. ni Embryo Whole IPLB-TN-R' Rochford et aI., 1984

T. ni Embryo Whole BTI-TN4Bl, Granados et al., 1986 -TN5Bl,-TN5Cl, -TN5F2, -TN5F2P, -TN5F2A, -TN5G2Al, -TN5G2AIP, -TN5G3, and -TN5G33

Olethreutidae Carpocapsa (= Cydia) Embryo Whole CP-169 and -1268 Hink and Ellis, 1971

pomonella C. pomonella Embryo Whole IZD-Cp-3300 Naser et aI., 1984

Papilionidae

Papilio xuthus Pupa Ovary NIAS-PX-58 Mitsuhashi,1973 and -64

Plutellidae

PluteUa xylostella Embryo Whole PX-1187 Lee and Hou, 1992 Pyralidae

Amyelois trnsitella Pupa Ovary HCRL-ATO-I0 Hoffmann et aI., 1990 and -20

Chilo suppressalis* Larva Hemocyte CSI6-CmIV Mitsuhashi, 1967

Ostrinia nubilalis Embryo Whole UMC-OnE Trisyono et aI., 2000

Plodia interpunctella Larva Imaginal IAL-PID2 Lynn and disc Oberlander, 1983



Saturniidae Antheraea eucalypti Pupa Ovary Ae Grace, 1962

Antheraea pernyi Embryo Whole NISES-AnPe -426 Inoue et aI., 1991

Samia cynthia Pupa Hemocyte Chao and Ball, 1971

Sphingidae

Manduca sexta Embryo Whole MRRL-CH-1 and-2 Eide et aI., 1975

M.sexta Embryo Whole GV-1 Lan and Riddiford, 1997

Tortricidae

Adoxophyes orana Neonate larva Whole FTRS-AoL-1 and -2 Sato,1989 fasciata

Adoxophyes sp. Neonate larva Whole FTRS-AfL Sato,1989 (smaller tea tortrix)

Archippus Neonate larva Whole FTRS-AbL-81 Sato,1989 breviplicanus

Choristoneura Larva Whole IPRI-Cf-124 Sohi,1973b fumiferana

C. fumiferana Larva Midgut Baines et aI., 1994

Homona Neonate larva Whole FTRS-HmL-45 Sato,1989 magnanima

Hoshinoa Neonate larva Whole FTRS-HlL-1 Sato,1989 longicellana and -2

Pandennis heparana Neonate larva Whole FTRS-PhL Sato,1989

Orthoptera Locustidae Anacridium Embryo Whole LPC-Aa 98-19, Hernandez-Crespo

aegyptium LPC-Aa 98-20 et al. 2000

Melanoplus Embryo Whole UMN-MSE Tsang et al. 1981 sanguinipes -1178-2

Aracnoidea Acarina Boophilus microplus Embryo Whole TTC-256 Pudney et aI., 1973

B. microplus Embryo Whole Holman and Ronald, 1980

Dermacentor Embryo Whole RML-14 Bhat and Yunker, parumapertus 1977

D. parumapertus Embryo Whole RML-16 and -17 Yunker et al., 1981

Dermacentor Embryo Whole RML-15 and -16 Yunker et aI., 1980 variabilis

D. variabilis Embryo Whole RML-18,-19, Yunker et al., 1981 and -20

Haemaphysalis Adult Developing ATC-304 and Guru et al.,1976 obesa tissue -311

430 Appendix


Haemophysalis Adult Developing ATC-2811282 Guru et aI., 1976 spinigera tissue andATC-309

Rhipicephalus Adult Developing TTC-219, -243, Varma et al., 1975 appendiculatus tissue and -257

Rhipicephalus Adult Developing ATC-307 and -308 Guru et aI., 1976 sanguineus tissue

Mollusca Gastropoda

Biomphalaria Embryo Whole Hansen, 1975 glabrata

Helix aspersa Adult Foot Vago and Chastang, 1958

Bivalvia

Meretrix lusoria Adult Heart Chen and Wen, 1999

Prochordata

Tunicata Botryllus schlosseri Embryo Whole NIO-BSE-l and-3 Rinkevich and

Rabinowitz, 1994

Nematoda

Tyrenchida

Meloidogyne Female adult Whole MI-8C and -5G Manousis and incognita Ellar, 1990

Plathelminthes Cestoda Echinococcus Cyst Furuya, 1991

multilocularis Coelenterata

Hydrozoa Millepora dichotoma Colony Fragment NIO-MD-l Frank et aI., 1994

Anthozoa

Clathrarcia Colony Fragment NIO-CR-l Frank et aI., 1994 rubinoides

Plexaura A Colony Fragment NIO-PC Frank et aI., 1994

Stylophora pistillata Colony Fragment NIO-SPP-l Frank et aI., 1994 planula larva

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434 Appendix

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Appendix 3

Commercially Available Culture Medium Products and List of Suppliers

A Commercially Available Culture Medium Products

Name of media Used for Supplier

Chiu & Black medium Leafhoppers Sigma

D-22 Drosophila melanogaster Sigma

Express Five SFM Trichoplusia ni Gibeo

Grace medium Most insects Sigma, Gibco

IPL-41 Most insects Sigma, Gibco

L-15 with 10% TPB Blood-sucking insects Sigma

L-15 + MEM (1: 1) Hank's Ticks Sigma salts + 10% TPB

MEM + Earl's salts and Aedes albopictus C6/36 Sigma non-essential amino acids clone

RPMI-1640 Echinococcus multilocularis Flow Laboratories Mitsuhashi and Most insects Sigma

Maramorosch medium

Schield and Sang medium M3 D. melanogaster Sigma Schneider medium Dipteran insects Sigma, Gibco

SF-900 II SFM Spodoptera frugiperda Gibco

TC-100 Lepidopteran insects Sigma, Gibeo

TNM-FH Lepidopteran insects Sigma

SFM, serum free medium; TPB, triptose phosphate broth; MEM, minimal essential medium.

B List of Suppliers

Agfa: Geneva, Switzerland American Ins.: Silver Spring, MD, USA American National Can Co.: Menasha, WI, USA American Type Culture Collection: Rockville, MD, USA Amicon Corp.: Danvers, MA, USA Aquarium Systems: Eastlake, OH, USA Arthur H. Thomas: Philadelphia, PA, USA Becton-Dickinson Labware: Lincoln Park, N], USA Bel-Art: Pequannock, N], USA Belleo Glass: Vineland, N], USA Biochrom: Berlin, Germany Bio-Rad: Richmond, CA, USA

436 Appendix

Boehringer-Mannheim Biochemicals: Indianapolis, IN, USA Brickman Instruments: Logan, UT, USA BTX, San Diego, CA, USA Calbiochem: La Jolla, CA, USA Clorox: Oakland, CA, USA Collaborative Research: Lexington, MA, USA Corning Medical, Corning Glass Works: Medfield, MA, USA Difco Laboratories: Detroit, MI, USA Dragon Chemical: Roanoke, VA, USA Drummond Sci, Co.: Broomall, PA, USA Falcon: Oxnard, CA, USA Flow Laboratories: Irvine, Scotland, UK FMC Bioproducts: Cambridge, MA, USA Fuji Photo Film: Tokyo, Japan Geisler Pet Products: New York, NY, USA Gentra Systems: Minneapolis, MN, USA Gibco-BRL: Grand Island, NY, USA Godo Shusei: Tokyo, Japan Greiner: Nuertingen, Germany Hampton Research: Laguna Niguel, CA, USA Hawaiian Marine Imports: Houston, TX, USA HEM Research: Rockville, MD. USA Humco-Sheffield: Norwich, NY, USA Ikemoto Rika: Tokyo, Japan Instant Ocean Aquarium System: Menton, OH, USA Jamarin Laboratory: Osaka, Japan KC Biological, Inc.: Lenexa, KS, USA Kontes Glass Co.: Vine land, NJ, USA Lab-Crest Scientific, Warminster, PA, USA LKB Instuments: Rockville, MD, USA Marco Specialty Steel, Inc.: Houston, TX, USA Marinus: Long Beach, CA, USA Merck: Darmstadt, Germany Microbiological Associates: Bethesda, MD, USA Millipore: Bedford, MA, USA Mitsubishi, Tokyo, Japan Nalgene (Nalge): Rochester, NY, USA New Brunswick Scientific: Edison, NJ, USA Nihon Nosan Kogyo, Yokohama, Japan Nikon, Tokyo, Japan Nipponseiyaku: Tokyo, Japan Nisseiken: Tokyo, Japan Nunc: Roskilde, Denmark Nutritional Biochemical: Cleveland, OH, USA Osi: Maurepas, France Oxoid: London, UK Pentex-Miles, Kankakee, IL. USA

Commercially Available Culture Medium Products and List of Suppliers 437

Perkin Elmer: Norwalk, CT, USA Pharmacia Fine Chemicals: Uppsala, Sweden Polaroid Corporation: Cambridge, MA,USA Razel Scientific Instruments, Inc., Stamford, CT, USA Riken Cell Bank: Ibaraki, Japan Robinson Laboratory: San Francisco, CA, USA Roche Chemicals: Nutley, NJ, USA Rohm and Haas: Philadelphia, PA, USA Roth: Karlsruhe, Germany Sartorius: G6ttingen, Germany Shin Etsu Chemical: Tokyo, Japan Sigma Chemical: St. Louis, MO, USA Societe Langlois: Niot, France Soden Chemicals: Toronto, Ontario, Canada Sovall: Manchester, UK Takara: Kyoto, Japan Tetko Inc.: Elmsford, NY, USA Toa Gosei Co. Ltd.: Tokyo, Japan VWR Scientific (Division ofUnivar): Columbus, OH, USA Verax Corporation: Lebanon, NH, USA Vestal Laboratories: St. Louis, MO, USA West Agro, Inc.: Kansascity, MO, USA Winthrop Laboratories: Surbiton, England; New York, NY, USA Woods End, Inc.: Weston, CT, USA Zoecon: Palo Alto, CA, USA

Subjuct Index

A

adult tissue - Arachnida (ticks) 124

Aedes aegypti 18 Aedes albopictus 76 Ahearn et al.'s saline 130 air-lift bioreactor 395 Amblyomma maculatum 128 amino acids 17 anesthesia (slugs) 283 Angiostrangylus cantonensis 205 Anomala costata 88, 89 Antheraea eucalypti 17 aphids 94 Apis mellifera 101, 103, 260 Arion subfuscus 283 Artemia 222, 275 Artemia nauplii 139 artificial seawater 143,150,156,188 aseptic rearing

- Drosophila 78, 241 Astacus leptodactylus 124, 269 Asterias amurensis 153, 281 Asterias rubens 154 Asterina gibbosa 281 Asterina pectinifera 281 Aurelia aurita 299 Australorbis glabratus 174 autoclave 7

B

Balanced salt solution (BSSO) (oysters) 401

Balanus amphitrite 139, 271, 275 barnacle organ culture medium IV 404 Basch and DiConza's medium C 404 bees 101 benzalkonium chloride 15 Biomphalaria glabrata 163 Blaberus craniifer 262

blastomeres - Mollusca (Gastropoda) 161

Blattella germanica 16,108 bleeding

- abalones 179 - coleopteran insects 89 - lepidopteran insects 59 - sea urchins 159

bodies - Mollusca (octopuses) 193

Bombyx mori 16,57,67,69, 255 Boophilus microplus 123 botryllid cell culture medium 405 Botryllus schlosseri 145,147,149,150 brain 64 buffered physiological solution with

antibiotics 401

c calcium alginate beads 397 Callinectes sapidus 270 Calliphora erythrocephala 251,257,259 capillary 308 capillary culture 127 cardiac tissue

- Mollusca (clams) 171 Cardisoma carnifex 272 Carlson's solution 27, 401 Carnoy's fixative 322 Cecil's medium modified by Heacox et al.

405 cell aspiration 308 cell counter 9 cell fusion

- Echinococcus cells 212 centrifuge 6 chemically defined media 21 Chernin's balanced salt solution 401 Chernin's nutrient medium 405 Chiarandini's saline solution 401

440 Subject Index

Chilo suppressalis 64, 65, 242, 255, 258

Chironomus ten tans 254 Chorthippus curtipennis 259 circulatory organ

- Lepidoptera 57 Clathrarcia rubrinoides 229 Clathrina 234 Clava leptostyla 300 cloning dish 317 coelomocytes

- Echinodermata (sea urchins) 158 - Echinodermata (starfish) 157

coenosarc - Coelenterata (Hydra) 224

collection of embryos (sponges) 235 colony formation 312, 314 corpora allata

- Blattaria 112, 249 corpora cardiaca

- Blattaria 112, 249 Cp medium 406 crab integument medium I and II 406 crab saline 401 Crassostrea gigas 176 Crassostrea virginica 176, 180 crayfish integument medium 406 cryopreservation 41 CSM-2F medium 406 Culex pipiens molestus 77 culture conditions 23 culture plate insert 319 culture vessels 13 cyst cells

- Platyhelminthes (Echinococcus) 209

cyst collection - (Echinococcus) 209

D

dechorionated egg culture (silkworms) 263

dechorionation - Drosophila 78 - silkworms 264

deep freezer 6 density gradient for separation of

- Hydra cells 228

- Echinococcus cell type 212 - neuronal cells (Drosophila) 82

Dermacentor andersoni 128 developer (photography) 353 dielectrophoresis 372 differentiation 53 digestion of mucus

- Mollusca(Bivalvia) 166 Diploptera punctata 250 dispase 35 dissecting microscope 6 dissecting tray 12 dissociation

- (sponge cells) 231,233 dissolution of virus inclusion bodies

359 dissolved oxygen concentration (DO)

387 DNA Amplification Fingerprinting (DAF)

329 DNA extraction 332 DNA-calcium phosphate coprecipitation

379 dorsal vessels

- Orthoptera 117 Drosophila 75, 80, 83 Drosophila extract 80 Drosophila hydei 254 Drosophila melanogaster 19, 73, 80,

84, 240, 254, 262 dry oven 7 Dugesia dorotocephala 218, 295 Dugesia tigrina 216 Dulbecco's modified Eagle's medium

407 Durchon and Schaller's medium for

Annelida 407

E

Echinococcus granulosus 210 Echinococcus multilocularis 211 Eisenia foetida 287,288 electrofusion 371 electrophoresis 333 electroporation 382 elimination of Protozoa

- Hydra 221 embryogenesis 67

embryos - Arachnida (ticks) 121 - Blattaria 107 - Coleoptera 87 - Crustacea (crayfish) 123 - Diptera 73 - Echinodermata (sea urchins) 156 - Echinodermata (starfish) 153 - Hemiptera 91 - Hymenoptera 102, 104 - Lepidoptera 66 - Mollusca (snails) 162 - Orthoptera 118 - Porifera (sponges) 235 - Tunicata 148

endoparasitoids 104 energy source 17 enigmatic vesicles (Sipunculoidea) 290 eosin Y 339 erythrosin B 338 essential amino acids 17 essential vitamins 18 Eunicella stricta 224, 301 Euproctis subflava 65 explant culture 23 exponential phase 350 eye

- Crustacea (Limulus) 274

F

fat body - Hymenoptera 260 - Lepidoptera 54

fat body extract 52 fatty acids 19 FBS 19, 21 fibrin clot 312 filter 30 filtration system 8 fixative (photography) 353 foot

- Mollusca (slugs) 167 FP1 401 free cell culture 22

G

y-irradiation 104 Galleria mellonella 240, 255, 259

Subject Index 441

gem mules (sponges) 231 germ-free aseptic rearing

- Planaria 215 germinal cells

- Platyhelminthes (Cestoda) 211 Gey's salt solution 402 Giemsa solution 322 gills

- Mollusca (clams) 184 gonad cells

- Crustacea (prawns) 137 gonad

- Echinodermata (starfish) 281 - Mollusca (cuttlefish) 284 - Mollusca (slugs) 283 - Proto chordata 279

gorgonian tissue (Hydra) 301 Gottschewski's medium 407 Grace 20 Grace's medium 27, 407 graduated pipettes 11 granular hemocytes 60 grid micrometer 349 growth factors 20 gut (Lepidoptera) 51

H

Haliotis discus 183 Haliotis discus culture medium 408 Haliotis rufescens 183 Haliotis rufescens culture medium 409 Haliotis tuberculata 179 Hansen's M-182 medium Hansen's M-260 medium Hansen's S-301 medium Heacox et al.'s Medium G heart

- Lepidoptera 57

408 408

408 409

- Mollusca (oysters) 175 - Mollusca (snails) 173

Heliothis virescens 52, 54 Helix aspersa 191 Helix pomatia 186 hemagglutinating virus of Japan (HVJ)

369 hemocytes

- Arachnida (ticks) 126 - Blattaria 108

442 Subject Index

- Coleoptera 88 - Diptera 84 - Lepidoptera 59 - Mollusca (abalones) 179 - Mollusca (oysters) 180 - Tunicata 145

hemocytometer 341 hemocytopoietic organ

- Lepidoptera 61 hemolymph 19 hepatopancreas

- Crustacea (shrimps) 130 Heremissenda crassicornis 188 Hirudo medicinalis 199, 288 Homona magnanima 69 honeybee medium 409 Hyalomma rufipes 123 hybrid-cell 37 Hydra (feeding) 223 Hydra culture solution 402 Hydra growth medium 409 Hydra littoralis 221 Hydra oligactis 300 Hydra pseudoligactis 300 Hydra viridis 300 Hymenolepis diminuta 296 Hyphantria cunea 49 hypotonic KCI solution 321

I

Ilyanassa obsoleta 161 imaginal disc

- Diptera 78, 240 - Lepidoptera 239

imaginal organ - leafhoppers 97

in vivo culture - Drosophila 265

Incilaria frukstorferi 167 incubator 5 inorganic salts 16 insect hormones 19 insect Ringer's solutions 402 integument

- Annelida (earthworms) 288 - Crustacea (crabs) 270 - Crustacea (crayfish) 269 - Lepidoptera 242

interstitial cell

- coelenterata (Hydra) 221 invertebrate Ringer's solution 402 inverted microscope 6 IPL-41 medium 409 irradiation 316 isocitrate dehydrogenase 326 isoelectric focusing 328 isolation of colonies 313 isotonic buffer for Echinococcus 402 isozyme analysis 325

J jar fermenter 392

K

K-6 medium 410 Kuroda and Tamura's medium 410

L

L-15 medium 410 L-15B 410 lactalbumin hydrolysate 21, 25 laminar flow cabinet 3, 15 Laodelphax striatellus 93 larval tissue

- leaf hoppers 95 Laverdure's medium 411 Laverdure's physiological solution 402 leafhoppers 91 leech Ringer's solution 402 Leloup's medium 1 411 Leucophaea maderae 262 Limulus 274, 277

- organ culture medium 411 lipofectin 40 lipofection 381 liquid nitrogen freezer 7 lobster hemolymph 133 Locke's salt solution 403 Lophogorgia sarmentosa 301 Lymantria dispar 52 lymphoid tissue

- Crustacea (shrimps) 131

M

Macrosteles fascifrons 97, 98 magnetic stirrer 10

malic enzyme 326 Malpighian tubes

- Lepidoptera 63 Mamestra brassicae 55,58,60,240,249 Manduca sexta 52, 242, 249 Mantle

- Crustacea (cirripeds) 271 - Mollusca (pearl oysters) 169 - Mollusca (slugs) 167 - Mollusca (Bivalvia) 165

Medium 1779 for Schistosoma 411 Medium 199 412 Medium 199 modified by Heacox et al.

412 Medium A-12 411 medium bottles 12 Medium C for snail cell culture 412 medium of Maiorella et al. 29 Medium X for Drosophila 413 Megoura crassicauda 95 melanization 20 Melanoplus sanguinipes 119, 120 Meloidogyne incognita 203, 293 Meretrix lusoria 184 metaphase arrest 321 methylcellulose 387 MGM-434 413 MGM-443 413 MGM-448 413 MGM-450 414 MGM-464 414 micro carrier 398 Microciona 234 micro elements 17 midgut glands

- Mollusca (Oncomelania) 189 midgut stem cells 52 Millepora dichotoma 229 Mizuchopecten yessoensis 183 MM medium 25, 26, 414 modified Alsever's anti-coagulant

solution 403 Molgula manhattensis 279 Molluscan balanced salt solution

(MMBSS) 403 MTT 343 multiplicity of infection (MOl) 361 mutation 376 Mycale contarenii 303

Subject Index 443

N

Na/K ratios 16 Necco's physiological solution 403 Negombata magnifica 235 neoblast

- Platyhelminthes (Planaria) 217 Neodiprion lecontei 102 neonate larvae

- Diptera 75 - Lepidoptera 69

neoplastic hemocyte 84 Nephotettix cincticeps 93 Nereis balanced salt solution 403 Nereis diversicolor 289 Nereis pelagica 289 Nereis virens 197 Nervous Tissue

- Annelida (leeches) 198 - Blattaria 109, 243, 246 - Diptera 81, 251 - Lepidoptera 64, 248 - Mollusca (Nudibranches) 188 - Mollusca (snails) 186

Neurosecretory Cells - Blattaria 111

nigrosin 339 Nudibranch culture medium 414

o Octopus physiological solution 403 ocular grid micrometer 342 oenocytoids 60 Ogura's medium 414 Oka organ 131, 132 oncogenes 39 Oncomelania culture medium 414 Oncomelania hupensis hupensis 190 oocytes

- Annelida (Nereis) 197 organic acids 19 Ornithodoros coriaceus 127 osmotic pressure 16 Ovary

- Coleoptera 259 - Crustacea (cirripeds) 138, 275 - Crustacea (crabs) 276 - Diptera 77 - Hemiptera 94

444 Subject Index

- Hymenoptera 101, 260 - Lepidoptera 47 - Nematoda 205

overlay medium 364 oviposition (ticks) 121 Ovotestis

- Mollusca (snails) 190

P

Pachygrapsus crassipes 272 packed-bed reactor 389 Palaemon serratus 130 palleal bud

- Tunicata 149 pancreatin 34 Panulirus homarus 133 Panulirus marginatus 272 Papilio xuthus 62, 63 parapodia

- Annelida (Nereis) 289 passage 36 Pasteur pipette 11, 15 Patiria miniata 158 PCR 331, 333 pearl chains 372 Penaeus japonicus 134 Penaeus monodon 131, 137, 138 Penaeus penicillantus 134 Penaeus stylirostris 132, 135 Penaeus vannamei 132 perfusion culture 393 perfusion module 393 Perinereis cultrifera 289 Periplaneta americana 18, 109, 110,

111, 113, 244, 248, 250 Pf-35 medium 415 pharyngeal tissue

- Tuniata 143 pheromone glands

- Lepidoptera 255 Philosamia cynthia 18 phosphoglucoisomerase 328 phosphoglucomutase 326 phosphoglucose isomerase 326 physiological solution 48 Pinctada fucata 167, 181 Pinctada fucata martensii 169 Pinctada margaritifera 167, 181

pipetting aids 12 Pisaster ochraceus segnis 158 planarian cell culture medium 415 planarian salt solution 403 planthoppers 93 plaque forming units (PFU) 363 plasmatocytes 60 Platynereis dumerilii 289 Plexaura A 229 Plodia interpunctella 240 Pluronic (polyol) F-68 30,387,391,395 polybrene-DMSO 380 polyethylene glycol (PEG) 370 Polymastia 234 porous carrier 399 prefiltration 31 preimaginal body

- Arachnida (ticks) 128 programmable freezer 6 prohemocytes 60 protein contents 344 protein products 19 prothoracic gland

- Lepidoptera 252 proventriculus

- Diptera 256 Pseudaletia separata 252

R

RAPD-PCR 331 Rasmont medium M 415 regenerating tissue

- Blattaria 261 regeneration blastema (Planaria) 295 replicate culture 347, 368 Rhipicephalus appendiculatus 123 Rhipicephalus sanguineus 128 Rinaldini's (salt) solution 33, 403 Ringer's salt solution 403 RML-375 medium 415 roller bottles 388 Ruditapes decussatus 184

S

S-20 medium 415 SA solution 404 salivary gland

- Diptera 253

Sarcophaga peregrina 18 Schistosoma mansoni 213 Schistosoma salt solution 404 scolex (cestodes) 296 Sengel's medium 416 separation of endoderm (Hydra) 299 separation of hypodermis (crayfish)

269 Sepia officinalis 284 septa

- Coelenterata (sea anemones) 226 serum-free media 21 SH-7 medium 416 shaking flask 390 shear force 387 shell opener 169 Shields and Sang's Medium 416

- modified by Poels 416 - modified by Sinha and Lakhotia

417 shrimp tissue culture medium (STCM)

132 silk gland

- Lepidoptera 254 Sipunculus nudus 291 sitting drops

- for cloning 311 snail organ culture medium 417 snail physiological saline solution 404 spermatocysts 50 spherule cells 60 spinner culture 390 Spisula solidissima 172 split-flow air-lift bioreactor 396 Spodoptera frugiperda 17 sponge Larva 303 spontaneous dissociation (Hydra) 228 stalk (Hydra) 300 sterilization

- abalones 183 - Mollusca (Bivalvia) 166 - Mollusca (oyster mantle) 170 - Mollusca{ clam cardiac tissue) 172 - Planaria 215 - root knot nematodes 203 - snail eggs 163

sterols 18 Strongylocentrotus franciscanus 159

Subject Index 445

Strongylocentrotus purpuratus 156,159 Styela clava 143 Stylophora pistillata 229 subculture 36 SV-40 large tumor antigen 40

T TAM-selection medium 374 TC-199 417 TC-199MK 417 TC-yeastolate 21, 27 TCEDso 368 TCIDso 361 Teichaxinella morchella 234 Tenebrio molitor 259 testis

- Lepidoptera 50 - Lepidoptera, Diptera, Orthoptera

257 testicular cells

- Crustacea 135 Thalamita crenata 276 thraustochytrids 166 tissue extract 20

- shrimp head 133 - shrimp muscle 133

Toxorhynchites amboinensis 76 trace metals 17 Trichogramma pretiosum 105 Trichoplusia ni 57 trochophore larvae (abalones) 183 trypan blue 337 trypsin 33 trypsinization (Hydra) 223 ts mutants 375 TTP medium 417 Tubularia crocea 225 Tubularia larynx 225 tumor

- Diptera 262 Tween-80 30 Tyrode's salt solution 404

U

ultra low-temperature freezer 7 urns

- Sipunculoidea 290

446 Subject Index

V

van Harreveld's saline 404 veligerlarvae

- (pearl oysters) 181 vesicles 56, 79, 108, 120 vitamins 18

W

water purification apparatus 9 water-insoluble substances 29 wing discs

- Lepidoptera 56 Wolff et al:s medium C 417 WS washing solution 404

Wyatt 20 Wyatt's medium 418

X

X-organ - Crustacea 272

y

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Composition of Salt Solutions and Culture Media978-4-431-67875...Composition of Salt Solutions and...

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Transcript of Composition of Salt Solutions and Culture Media978-4-431-67875...Composition of Salt Solutions and...