Edition 02/2014 - MCH LAB• The MSDS for GM501 Wash is available upon request ... precautions...

Edition 02/2014

Micropipettes

Oocyte Handling Media 1

Culture Media 2

Sperm Processing Media 3

Cryopreservation Media 4

Miscellaneous 7

Micropipettes 5

Denudation-Tips 6

GYNEMED GmbH & Co. KG ○ Lübecker Str. 9 ○ 23738 Lensahn ○ Germany phone: +49 (0) 4363 903 290 ○ fax: +49 (0) 4363 903 2919

http://www.gynemed.de ○ e-mail: [email protected]

Oocyte Handling Media 1

GM501 Wash

Product code: 1 x 20 ml 4 GM 501W-201 x 50 ml 4 GM 501W-501 x 500 ml 4 GM 501W-500

GM501 Wash is a ready-to-use medium designed for wa-shing procedures of human oocytes and embryos and any short-term handling procedures outside the incubator like washing after Hyaluronidase treatment (denudation), ICSI, polar body or blastomere biopsy and other.

The intended users are IVF professionals (lab technicians, embryologists or medical doctors).

Composition:• NaCl, KCl, KH2PO4, MgSO4

.7H2O, NaHCO3, CaCl2.2H2O,

D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-essential and essential Amino Acids, HEPES, Human serum albumin (5.00 g/liter).

Instructions for use:• GM501 Wash must be equilibrated over night in a hu-

midified CO2-incubator (at 5-7 % CO2, 37°C).

Product specifications and quality control: • All raw materials are of highest available purity (Euro-

pean Pharmacopoeia and/or USP standard), if appli-cable.

• A certificate of analysis is available for each batch upon request from our website with respective lot number.

• The MSDS for GM501 Wash is available upon request and can also be downloaded from our website.

• GM501 Wash is manufactured and tested according to the following specifications:

pH (at 37°C, 6 % CO2) 7.20-7.50

Osmolality (mOsm/kg) 270-290

Sterility sterile - SAL10-3 (Sterility Assurance Level)

Endotoxins (EU/ml) < 0.25

MEA (Blastocysts after 96h in %)

≥ 80after 60 min exposure

(zygote stage)

Precautions and warnings:• Standard measures to prevent infections resulting

from the use of medicinal products prepared from human blood or plasma include selection of donors, screening of individual donations and plasma pools for specific markers of infection and the inclusion of effective manufacturing steps for the inactivati-on/removal of viruses. Despite this, when medicinal products prepared from human blood or plasma are administered, the possibility of transmitting infective agents cannot be totally excluded. This also applies to unknown or emerging viruses and other patho-gens. There are no reports of proven virus transmis-sions with albumin manufactured to European Phar-macopoeia specifications by established processes.

• Therefore, handle all specimens as if capable of transmitting HIV or hepatitis. Always wear protective clothing when handling specimens.

• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contami-nation.

• Only for the intended use.

Pre-use checks:• Do not use the product if bottle, seal of the container

or package is opened or defect when the product is delivered.

• Do not use the product if it becomes discoloured, cloudy or shows any evidence of microbial contami-nation.

Storage instructions and stability:• Shelf life is 6 months from time of manufacture.• Store between 2-8°C. • Do not freeze before use.• Keep away from (sun) light.• The product can be used safely up to 7 days after

opening, when sterile conditions are maintained.• Do not use after expiry date.

0344



GM501 Wash with Phenolred and Gentamicin

Product code: 1 x 20 ml 4 GM 501W+PR-20

1 x 50 ml 4 GM 501W+PR-50

GM501 Wash with Phenolred and Gentamicin is a ready-to-use medium designed for washing procedures of hu-man oocytes and embryos and any short-term handling procedures outside the incubator like washing after Hy-aluronidase treatment (denudation), ICSI, polar body or blastomere biopsy and other.




D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-essential and essential Amino Acids, HEPES, Human serum albumin (5.00 g/liter), Gentamicin (10 mg/liter), Phenolred.

Instructions for use:• GM501 Wash with Phenolred and Gentamicin must

be equilibrated over night in a humidified CO2-incu-bator (at 5-7 % CO2, 37°C).




• The MSDS for GM501 Wash with Phenolred and Gen-tamicin is available upon request and can also be downloaded from our website.

• GM501 Wash with Phenolred and Gentamicin is manu-factured and tested according to the following speci-fications:

pH (at 37°C, 6 % CO2) 7.20-7.50





≥ 80after 60 min exposure

(zygote stage)

0344

Precautions and warnings:• GM501 Wash with Phenolred and Gentamicin con-

tains the antibiotic Genta micin Sulfate. Appropriate precautions should be taken to ensure that the pati-ent is not sensitized to this antibiotic.

• Standard measures to prevent infections resulting from the use of medicinal products prepared from human blood or plasma include selection of donors, screening of individual donations and plasma pools for specific markers of infection and the inclusion of effective manufacturing steps for the inactivati-on/removal of viruses. Despite this, when medicinal products prepared from human blood or plasma are administered, the possibility of transmitting infective agents cannot be totally excluded. This also applies to unknown or emerging viruses and other patho-gens. There are no reports of proven virus transmis-sions with albumin manufactured to European Phar-macopoeia specifications by established processes.


• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contami-nation, even when GM501 Wash medium contains Gentamicin.







Oocyte Handling Media1



Oocyte Handling Media Oocyte Handling Media 1

GM501 Hyaluronidase

Product code: 1 x 10 ml 4 HY 0010

5 x 1 ml 4 HY 0001-5

GM501 Hyaluronidase is a ready-to-use solution desi-gned to digest the extra cellular matrix of hyaluronic acid in the cumulus oocyte complex.


Composition:• 80 IU/ml pharmacentical grade hyaluronidase solved

in HEPES-buffered medium.

Instructions for use:• GM501 Hyaluronidase contains HEPES; no CO2-incu-

bation is required, just warm it up to 37°C.• Depending on the dishes, 100 µl (microdrops) or

400-600 µl (center-/4-well) of GM501 Hyaluronidase should be used for denudation. The dishes should be covered with Mineral Oil (e.g. GM501 Mineral Oil see pg. 13). Place the oocytes in the GM501 Hyaluronidase (up to max. 5 oocytes) for about 30 seconds. Using a fine pipette, transfer the partially denuded oocytes to the first washing droplet. Remove the corona by pipetting the oocytes (e.g. Denudation-Tips see pg. 34-35). Use the other droplets to further wash the denuded oocyte.

Product specifications and quality control:• All raw materials are of highest available purity (Euro-



• The MSDS for GM501 Hyaluronidase is available upon request and can also be downloaded from our website.

• GM501 Hyaluronidase is manufactured according to the following specifications:

pH (at 37°C) 7.20-7.60














Due to national legislation is this product not available in all countries!


≥ 80



Oocyte Handling Media1

GM501 Hyaluronidase: Recommended application

Preparation:We recommend the following products from the GM501 MediaLine:• GM501 Hyaluronidase• GM501 Cult• GM501 Wash• GM501 Mineral Oil

Oocyte Retrieval:• Prepare all needed dishes with GM501 Wash. The

GM501 Wash must be equilibrated over night in a hu-midified CO2-incubator at 37°C prior to use.

• Transfer the follicles by aspirating them into empty dishes. Identify the oocytes and immediately remove them from the follicle fluid. Rinse them in equilibrated GM501 Wash or GM501 Cult Media and transfer them to equilibrated culture dishes (ICSI) or fertilization dis-hes with GM501 Cult Media until denudation or inse-mination for IVF.

Preparation for denudation of fresh oocytes:Preparation of the 4-well dishes • For up to 10 oocytes one 4-well-dish is necessary to

prepare the “Hya-dish”. • First fill wells 2 to 4 with 400 µl GM501 Wash medi-

um and equilibrate the dish over night in a humidi-fied CO2-incubator at 37°C. Cover the filled wells with GM501 Mineral Oil.

• Warm the GM501 Hyaluronidase and fill the first well with 400 µl warmed GM501 Hyaluronidase.

• Now using a serological pipette (e.g. MEA-tested Ep-pendorf pipettes) for further procedures.

Preparation of the 60 mm dishes • For up to 10 oocytes one 60 mm Petri dish is recom-

mended to prepare the “Hya-dish”. Each dish con-tains 7 drops, 80 µl of volume each.

• There are 6 outer drops and one central drop. One of the outer drops is marked by a circle at the bottom of the dish using a permanet marker.

• First fill the dish with 6 drops of 80 µl GM501 Wash and equilibrate the dish over night in a humidified CO2-incubator at 37°C. Cover the drops with GM501 Mineral Oil.

• Warm the GM501 Hyaluronidase and fill the dish with one drop of 80 µl warmed GM501 Hyaluronidase.

• Now using a serological pipette (e.g. MEA-tested Ep-pendorf pipettes) for further procedures.

400 µlGM501

Hyaluro-nidase

400 µlGM501 Wash

400 µl GM501 Wash

400 µlGM501 Wash

80 µl GM501 WashMedium/drop

80 µl hyaluronidase- solution



Oocyte Handling Media Oocyte Handling Media 1

GM501 Hyaluronidase: Recommended application

Denudation Procedure with Microdrop-Dishes

1. Take all the dish(es) containing the fresh oocytes out of the incubator and place them under a stereomicro-scope.

2. Pipette up to 10 oocytes into the central drop of the “Hya-dish“ using the Eppendorf-pipette (100 µl-tip adjusted to 50 µl)

3. If there are more than 10 oocytes, put the supernu-merary back into the incubator.

4. Transfer 5 oocytes to the “Hya-drop” of the “Hya-dish”.

5. Immediately aspirate and blow them out repeatedly into the “Hya-drop” (5 to 10 times).

6. The cumulus will decompose into single cells and the oocytes still surrounded by corona cells will get vis-ible.

Attention: The oocytes should not be in the Hya-solution longer than 30 seconds!

7. Pick up the oocytes using the denudation pipette (inner diameter 125–155 µm, e.g. Denudation-Tips see pp. 34-35) and transfer them to the drop con-taining GM501 Wash medium at the right side of the “Hya-drop”.

8. Aspirate the oocytes and blow them back and forth repeatedly.

9. Pick up all oocytes and transfer them to the next drop to the right. Again aspirate and blow them out repeat-edly until nearly all corona cells are removed.

10. Transfer the oocytes to the next drop to the right and leave them in the drop.

11. Now take the Eppendorf-pipette again and transfer the next 5 oocytes that are still in the central drop to the “Hya-drop”.

12. Repeat steps 4 to 10 with these oocytes.

13. When all oocytes are collected in GM501 Wash drop three and are free of cumulus and corona cells wash them in the last two drops as described and transfer them back to a culture dish with GM501 Cult Media for incubation until ICSI or directly to the ICSI-dish.



Culture MediaCulture Media2

GM501 Cult

Product code: 1 x 20 ml 4 GM 501H-20

1 x 50 ml 4 GM 501H-50

GM501 Cult is a ready-to-use bicarbonate-buffered cul-ture media, designed for fertilisation and for human emb-ryo culture from day 1 to blastocyst stage. It can also be used for embryo transfer.




D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-es-sential and essential Amino Acids, Human serum albu-min (10.00 g/liter).

Instructions for use:• GM501 Cult must be equilibrated over night in a hu-

midified CO2-incubator (at 5-7 % CO2, 37°C).• The medium is designed for embryo culture under oil

as well as in open systems.




• The MSDS for GM501 Cult is available upon request and can also be downloaded from our website.

• GM501 Cult is manufactured and tested according to the following specifications:

pH (at 37°C, 6 % CO2) 7.20-7.45


Sterility sterile -SAL10-3 (Sterility Assurance Level)



≥ 80(after 96h exposure from

zygote stage)

0344Precautions and warnings:• Standard measures to prevent infections resulting from

the use of medicinal products prepared from human blood or plasma include selection of donors, screening of individual donations and plasma pools for speci-fic markers of infection and the inclusion of effective manufacturing steps for the inactivation/removal of viruses. Despite this, when medicinal products pre-pared from human blood or plasma are administered, the possibility of transmitting infective agents cannot be totally excluded. This also applies to unknown or emerging viruses and other pathogens. There are no reports of proven virus transmissions with albumin ma-nufactured to European Pharmacopoeia specifications by established processes.


• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contamination.




• Do not use the product if it becomes discoloured, clou-dy or shows any evidence of microbial contamination.



References:1. Eue I., Schwahn E., Merzenich M., Häberlin F. (2007): First

Clinical Experience with GM501 - a New KSOMAA based Embryo Culture Medium. Annual meeting of German IVF-Groups, Kiel.

2. Weiss D., Schneider U. (2006): In vitro culture of human em-bryos up to day five in simplex optimized medium GM501. Gynemedia.



Culture Media 2Culture Media

GM501 Cult with Gentamicin

Product code: 1 x 20 ml 4 GM 501H+G-20

1 x 50 ml 4 GM 501H+G-50

GM501 Cult with Gentamicin is a ready-to-use bicarbo-nate-buffered culture media, designed for fertilisation and for human embryo culture from day 1 to blastocyst stage. It can also be used for embryo transfer.



.7H2O, NaHCO3, CaCl2

.2H2O, D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutami-ne, Water, non-essential and essential Amino Acids, Human serum albumin (10.00 g/liter), Gentamicin (10.00 mg/liter).

Instructions for use:• GM501 Cult with Gentamicin must be equilibrated

over night in a humidified CO2-incubator (at 5-7 % CO2, 37°C).

• The medium is designed for embryo culture under oil as well as in open systems.




• The MSDS for GM501 Cult with Gentamicin is availa-ble upon request and can also be downloaded from our website.

• GM501 Cult with Gentamicin is manufactured and tested according to the following specifications:

0344

pH (at 37°C, 6 % CO2) 7.20-7.45






zygote stage)

Precautions and warnings:• GM501 Cult with Gentamicin contains the antibiotic

Genta micin Sulfate. Appropriate precautions should be taken to ensure that the patient is not sensitized to this antibiotic.

• Standard measures to prevent infections resulting from the use of medicinal products prepared from human blood or plasma include selection of donors, screening of individual donations and plasma pools for speci-fic markers of infection and the inclusion of effective manufacturing steps for the inactivation/removal of viruses. Despite this, when medicinal products pre-pared from human blood or plasma are administered, the possibility of transmitting infective agents cannot be totally excluded. This also applies to unknown or emerging viruses and other pathogens. There are no reports of proven virus transmissions with albumin ma-nufactured to European Pharmacopoeia specifications by established processes.


• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contamination, even when GM501 Cult medium contains Gentamicin.







References:1. Kemeter P., Hajek J., Feichtinger W. (2011): Eine prospek-

tiv-randomisierte Studie zum Vergleich zweier Embryo-Kultursysteme nach IVF/ICSI: Sequential media in 5 % O2-Atmosphäre und Single medium in 21 % O2-Atmosphäre. J Gynäkol Endokrinol 21: 16-21.

2. Campo R., Binda M.M., Van Kerkhoven G., Frederickx V., Serneels A., Roziers P., Lopes A.S., Gordts Sy., Puttemans P., Gordts S. (2010): Critical reappraisal of embryo quality as a predictive parameter for pregnancy outcome: a pilot study. F, V & V in ObGyn 2: 289-295.




GM501 Cult with Gentamicin and Phenolred

Product code: 1 x 20 ml 4 GM 501H+PR+G-20

1 x 50 ml 4 GM 501H+PR+G-50

GM501 Cult with Gentamicin and Phenolred is a ready-to-use bicarbonate-buffered culture media, designed for fertilisation and for human embryo culture from day 1 to blastocyst stage. It can also be used for embryo transfer.




D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-essential and essential Amino Acids, Human serum albumin (10.00 g/liter), Gentamicin (10.00 mg/liter), Phenolred.

Instructions for use:• GM501 Cult with Gentamicin and Phenolred must be

equilibrated over night in a humidified CO2-incubator (at 5-7 % CO2, 37°C).

• The medium is designed for embryo culture under oil as well as in open systems.




• The MSDS for GM501 Cult with Gentamicin and Phe-nolred is available upon request and can also be down-loaded from our website.

• GM501 Cult with Gentamicin and Phenolred is manu-factured and tested according to the following specifi-cations:

Precautions and warnings:• GM501 Cult with Gentamicin and Phenolred contains

the antibiotic Genta micin Sulfate. Appropriate precau-tions should be taken to ensure that the patient is not sensitized to this antibiotic.

• Standard measures to prevent infections resulting from the use of medicinal products prepared from human blood or plasma include selection of donors, scree-ning of individual donations and plasma pools for spe-cific markers of infection and the inclusion of effective manufacturing steps for the inactivation/removal of viruses. Despite this, when medicinal products pre-pared from human blood or plasma are administered, the possibility of transmitting infective agents cannot be totally excluded. This also applies to unknown or emerging viruses and other pathogens. There are no reports of proven virus transmissions with albumin manufactured to European Pharmacopoeia specifica-tions by established processes.


• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contami-nation, even when GM501 Cult with Gentamicin and Phenolred medium contains Gentamicin.







References:1. Gruber I., Klein M. (2011): Embryo culture media for human

IVF: which possibilities exist? J Turkish-German Gynecol As-soc 12: 110-117.

2. Paternot G., Debrock S., D´Hooghe T.M., Spiessens C. (2010): Early embryo development in a sequential versus sin-gle medium: a randomized study. Reproductive Biology and Endocrinology 7: 83.

pH (at 37°C, 6 % CO2) 7.20-7.45






zygote stage)

0344




GM501 Basic

Product code: 1 x 50 ml 4 GM 501-50

GM501 Basic is a bicarbonate-buffered culture media, designed for fertilisation and for all embryo culture from day 1 to blastocyst stage. It can also be used for embryo transfer.




D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-es-sential and essential Amino Acids.

Instructions for use:Add human serum albumin prior to use (preparation of working solution).• GM501 Basic must be supplemented with human

serum albumin to a protein end concentration of 1 % (w/v). The use of 4 HSA-0005 (Gynemed) is strongly recommended. To supplement the 50 ml unit, keep 45 ml medium in the bottle and add 5 ml of 4 HSA-0005. Mix well.

• Embryo culture media must be equilibrated over night in a humidified CO2-incubator (at 5-7 % CO2, 37°C).

• GM501 Basic is designed for embryo culture under oil as well as in open systems.




• The MSDS for GM501 Basic is available upon request and can also be downloaded from our website.

• GM501 Basic is manufactured and tested according to the following specifications:

Precautions and warnings:• Handle all specimens as if capable of transmitting HIV

or hepatitis. Always wear protective clothing when handling specimens.






• Addition of human serum albumin by the customer is required for optimal culture conditions.



References:1. Ebert P., Szypajlo B., Tomalak K., Völklein K. (2009): Prospec-

tive comparison of two commercially available culture media under the provisions of the German embryo protection law. J Turkish-German Gynecol Assoc 10: 10-13

2. Eue I., Schwahn E., Merzenich M., Häberlin F. (2007): First Clinical Experience with GM501 - a New KSOMAA based Embryo Culture Medium. Annual meeting of German IVF-Groups, Kiel.

3. Weiss D., Schneider U. (2006): In vitro culture of human em-bryos up to day five in simplex optimized medium GM501. Gynemedia.

0482

pH (at 37°C, 6 % CO2) 7.20-7.40






zygote stage)



Culture Media2

GM501 Basic with Gentamicin

Product code: 1 x 50 ml 4 GM 501+G-50

GM501 Basic with Gentamicin is a bicarbonate-buffered culture media, designed for fertilisation and for all embryo culture from day 1 to blastocyst stage. It can also be used for embryo transfer.




D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non- essential and essential Amino Acids, Gentamicin (10.00 mg/liter).

Instructions for use:Add human serum albumin prior to use (preparation of working solution).• GM501 Basic with Gentamicin must be supple-

mented with human serum albumin to a protein end concentration of 1 % (w/v). The use of 4 HSA-0005 (Gynemed) is strongly recommended. To supplement the 50 ml unit, keep 45 ml medium in the bottle and add 5 ml of 4 HSA-0005. Mix well.

• Embryo culture media must be equilibrated over night in a humidified CO2-incubator (at 5-7 % CO2, 37°C).

• GM501 Basic with Gentamicin is designed for emb-ryo culture under oil as well as in open systems.

Product specifications and quality control: • All raw materials are of highest available purity (Eu-

ropean Pharmacopoeia and/or USP standard), if ap-plicable.


• The MSDS for GM501 Basic with Gentamicin is available upon request and can also be downloaded from our website.

• GM501 Basic with Gentamicin is manufactured and tested according to the following specifications:

Precautions and warnings:• GM501 Basic with Gentamicin contains the antibiotic

Genta micin Sulfate. Appropriate precautions should be taken to ensure that the patient is not sensitized to this antibiotic.

• Handle all specimens as if capable of transmitting HIV or hepatitis. Always wear protective clothing when handling specimens.

• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contamination, even when GM501 Basic with Gentamicin medium contains Gentamicin.





• Addition of human serum albumin by the customer is required for optimal culture conditions.



References:1. Ebert P., Szypajlo B., Tomalak K., Völklein K. (2009): Prospective

comparison of two commercially available culture media under the provisions of the German embryo protection law. J Turkish-German Gynecol Assoc 10: 10-13

2. Eue I., Schwahn E., Merzenich M., Häberlin F. (2007): First Cli-nical Experience with GM501 - a New KSOMAA based Embryo Culture Medium. Annual meeting of German IVF-Groups, Kiel.

3. Weiss D., Schneider U. (2006): In vitro culture of human emb-ryos up to day five in simplex optimized medium GM501. Gy-nemedia.

0482

pH (at 37°C, 6 % CO2) 7.20-7.40






zygote stage)








zygote stage)

GM501 Cult: Recommended application (IVF)Preparation:We recommend the following products from the GM501 MediaLine:• GM501 Cult or• GM501 Cult with Gentamicin or• GM501 Cult with Gentamicin and Phenolred• GM501 Mineral Oil

4-well-dish:• At day “-1” prepare a 4-well-fertilization-dish by fill-

ing each well with 400 µl of GM501 Cult Media. Cover the dish completely with GM501 Mineral Oil and incu-bate it in a CO2-atmosphere at 37°C.

• At day “0” add up to 6 oocytes per well for insemina-tion.

• Prepare a 4-well-culture-dish for day 1 in the same way like the fertilization dish.

• At day “1” partly remove the cumulus and granulosa cells from the inseminated oocytes and transfer the fertilized oocytes to the culture dish. Add up to 6 oo-cytes in one well for ongoing culture.

400 µlGM501

CultMedia

400 µlGM501

CultMedia

400 µlGM501

CultMedia

400 µlGM501

CultMedia

Day -1 Day 0 Day 1 Day 2 or 3

Preparation of fertilization dishes (IVF) using

GM501 Cult Media

Fertilization by IVF and culture in GM501 Cult

Media

Ongoing culture in GM501 Cult Media

Embryo transfer

Day -1 Day 0 Day 1 Day 2 Day 3 Day 4 Day 5

Preparation of fertilization dishes (IVF)

using GM501 Cult Media

Fertilization by IVF and

culture in GM501 Cult

Media

Ongoing culture in

GM501 Cult Media

Ongoing culture in

GM501 Cult Media.

Preparation of new dishes for day 3 if wanted

Ongoing culture in new

dishes with GM501 Cult

Media

Ongoing culture in

GM501 Cult Media

Embryo transfer

Simple „one-step“ culture:

Extended „one-step“ culture:

Microdrop dish:• At day “-1” prepare a microdrop-fertilization-dish

by setting several 80–100 µl drops in a 60 mm Petri-dish. Cover the dish completely with GM501 Mineral Oil and incubate it in a CO2-atmosphere at 37°C.

• At day “0” add up to 2 oocytes per drop for insemina-tion.

• Prepare a microdrop-culture-dish for day 1 in the same way like the fertilization dish.

• At day “1” partly remove the cumulus and granulosa cells from the inseminated oocytes and transfer the fertilized oocytes to the culture dish. Add up to 3 oo-cytes in one drop for ongoing culture. For single-cell culture 40–50 µl drops should be used.

40-50 µlGM501 Cult Media

(single-cell culture)


(group culture)

or




GM501 Cult: Recommended application (ICSI)Preparation:We recommend the following products from the GM501 MediaLine:• GM501 Cult or• GM501 Cult with Gentamicin or• GM501 Cult with Gentamicin and Phenolred or• GM501 Mineral Oil

4-well-dish:• At day “-1” prepare a 4-well-culture-dish by filling

each well with 400 µl of GM501 Cult Media.• Cover the dish completely with GM501 Mineral Oil

and incubate it in a CO2-atmosphere at 37°C.• Right after microinjection at day “0” add up to 6 oo-

cytes per well for ongoing culture.

400 µlGM501

CultMedia

400 µlGM501

Cult Media

400 µlGM501

CultMedia

400 µlGM501

CultMedia

Day -1 Day 0 Day 1 Day 2 or 3

Preparation of culture dishes (ICSI) using GM501 Cult Media

Fertilization by ICSI and culture in GM501 Cult

Media

Ongoing culture in GM501 Cult Media

Embryo transfer


Preparation of culture dishes

(ICSI) using GM501 Cult

Media

Fertilization by ICSI and culture in

GM501 Cult Media

Ongoing culture in

GM501 Cult Media

Ongoing culture in

GM501 Cult Media.




Media

Ongoing culture in

GM501 Cult Media

Embryo transfer

Simple „one-step“ culture:

Extended „one-step“ culture:

Microdrop dish:• At day “-1” prepare a microdrop-culture-dish by

setting several 80–100 µl drops in a 60 mm Petri dish. • Cover the dish completely with GM501 Mineral Oil

and incubate it in a CO2-atmosphere at 37°C.• Right after microinjection at day “0” add up to 3 oo-

cytes per drop for ongoing culture.• For single-cell culture 40–50 µl drops should be used.


(single-cell culture)


(group culture)

or





Preparation of culture dishes

(ICSI) using GM501 Cult

Media

Fertilization by ICSI and culture in

GM501 Cult Media

Ongoing culture in

GM501 Cult Media

Ongoing culture in

GM501 Cult Media.




Media

Ongoing culture in

GM501 Cult Media

Embryo transfer

GM501 Mineral Oil

GM501 Mineral Oil is a ready-to-use mineral oil to reduce evaporation and to stabilize the pH during IVF, ICSI and related artificial reproductive techniques.


Composition:• GM501 Mineral Oil is a pre-washed light density mi-

neral oil (d = 0.83-0.86 g/ml).

Instructions for use:• GM501 Mineral Oil is a pre-washed ready-to-use oil,

so no further preparations are necessary.• Simply the GM501 Mineral Oil has to be pre-incuba-

ted in a incubator at 37°C for 5 hours before use (with lid closed).

• Overlay the Cell culture medium with GM501 Mineral Oil until it is completely covered.

Product specifications and quality control:• Pharmaceutical grade mineral (paraffin) oil.• A certificate of analysis is available for each batch

upon request from our website with respective lot number.

• The MSDS for GM501 Mineral Oil is available upon re-quest and can also be downloaded from our website.

• GM501 Mineral Oil is manufactured according to the following specifications:

Product code: 1 x 100 ml 4 MO 0100

1 x 500 ml 4 MO 0500

Density (g/ml) 0.83-0.86

Viscosity (cP at 30°C) < 30



POV (mEq/Kg)(Peroxide value)

< 0.1


≥ 80




• Only for the intended use.• The oil may appear cloudy if removed from the incu-

bator and allowed to stand at room temperature for extended periods of time. Such cloudiness usually disappears if the oil is re-equilibrated by replacing it in the incubator for several hours, if it does not disap-pear discard the product.




Storage instructions and stability:• Shelf life is 18 months from time of manu facture.• Store between 15-25°C. • Keep the product in the original packaging or shield

from light while not in use.• Do not freeze before use.• Keep away from (sun) light.• The product can be used safely for at least 28 days

after opening the container when sterile conditions are maintained and the products are stored at room temperature.

• Do not use after expiry date.

0482



Sperm Processing MediaSperm Processing Media3

GM501 SpermAir

Product code: 1 x 20 ml 4 GM 501AIR+G+PR-20

1 x 50 ml 4 GM 501AIR+G+PR-50

GM501 SpermAir is a ready-to-use medium designed for all human sperm-preparation, sperm-washing and swim-up techniques like density-gradient centrifugation as well as for testicular tissue.


Composition:• NaCl, KCI, KH2PO4, MgSO4


D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, water, non-es-sential and essential Amino Acids, 21mM HEPES, Hu-man serum albumin (5.00 g/liter), Gentamicin (10.00 mg/liter), Phenolred.

Instructions for use:• Do not equilibrate GM501 SpermAir in a CO2-incu-

bator, just warm it up to 37°C. GM501 SpermAir is HEPES-buffered. Incubation in a CO2-incubator will lower the pH.




• The MSDS for GM501 SpermAir is available upon re-quest and can also be downloaded from our website.

• GM501 SpermAir is manufactured and tested accord-ing to the following specifications:

pH (at 37°C) 7.20-7.50




Precautions and warnings:• GM501 SpermAir contains the antibiotic Genta micin

Sulfate. Appropriate precautions should be taken to ensure that the patient is not sensitized to this anti-biotic.



• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contami-nation, even when GM501 SpermAir medium con-tains Gentamicin.







0344

MEA (Blastocysts in %)

≥ 80(after 5 days of culture in the product covered by mineral oil, starting

from zygote stage)



Sperm Processing Media 3Sperm Processing Media

GM501 SpermActive is a ready-to-use medium designed for all sperm-preparation, sperm-washing and swim-up techniques like density-gradient centrifugation as well as for testicular tissue.




D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-es-sential and essential Amino Acids, HEPES, Human se-rum albumin (5.00 g/liter), Gentamicin (10.00 mg/liter), Phenolred.

Instructions for use:• GM501 SpermActive must be equilibrated over night

in a humidified CO2-incubator (at 5-7 % CO2, 37°C).




• The MSDS for GM501 SpermActive is available upon request and can also be downloaded from our website.

• GM501 SpermActive is manufactured and tested ac-cording to the following specifications:

GM501 SpermActive

Product code: 1 x 20 ml 4 GM 501SA-020

1 x 50 ml 4 GM 501SA-050

pH (at 37°C, 6 % CO2) 7.20-7.50





≥ 80after 60 min of exposure

(zygote stage)

0344

Precautions and warnings:• GM501 SpermActive contains the antibiotic Genta-

micin Sulfate. Appropriate precautions should be ta-ken to ensure that the patient is not sensitized to this antibiotic.



• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contami-nation, even when GM501 SpermActive medium con-tains Gentamicin.









Sperm Processing Media3

Recommended application: Mini-Swim-up

Preparation:We recommend one of the products from the GM501 MediaLine:• GM501 SpermAir or• GM501 SpermActive

Take the semen sample, put it for at least 20 min. at 37ºC on a warming plate or a warming area until it is liquefied.

3.

4.

6.

1.

2.

7.

8.

5.

9.

⟳Mix 1-3 ml liquefied ejaculate with 4.5 ml equilibrated GM501 Sperm-Active or warmed GM501 SpermAir in a conical tube.

Centrifuge the tube at 300-400 g for 10 minutes.

Aspirate the supernatant without dispersing the pellet and discard it.

Resuspend the pellet with 1 ml equilibrated GM501 SpermActive or warmed GM501 SpermAir.

Fill the suspension in a 1.5 ml Ep-pendorf tube.


⟳


Resuspend the pellet with 0.5 ml equilibrated GM501 SpermActive or warmed GM501 SpermAir.


⟳Aspirate the supernatant without dispersing the pellet and discard it.

Overlay the pellet with 25-50 µl equilibrated GM501 SpermActive or warmed GM501 SpermAir.

Close the top and stich a hole in the lid by using a sterile cannula to guarantee the contact with CO2 and O2.

Put the tube for 1 hour into the CO2-incubator if you use GM501 SpermActive or in a heating cabi-net if you use GM501 SpermAir. Use the sperms in the supernatant for ICSI.




Recommended application: Direct Swim-up

To prepare the swim-up tubes, transfer 2 ml medium into a new tube.

Underlay gently 1 ml of liquefied semen.

1.

2. Place the tube in the CO2-incubator (GM501 SpermActive) or heating cabinet (GM501 SpermAir) for 1 hour with not firmly closed top.

Aspirate the upper media layer containing the motile sperms with-out dispersing the native ejaculate and fill it into a new conical centri-fuge tube.

3.

4. Centrifuge the tube at 300-400 g for 6 minutes.


Resuspend the pellet with 1 ml equilibrated GM501 SpermActive or warmed GM501 SpermAir me-dium.

5.

6. Centrifuge the tube at 300-400 g for 6 minutes.

Repeat step 5.

Aspirate the supernatant and dis-card it.

For IVF resuspend the pellet in 0.1–1 ml equilibrated culture medium (e.g. GM501 Culture Media) to the desired sperm concentration, for ICSI or Insemination resuspend it in GM501 SpermActive or GM501 SpermAir.

7.

8.



⟳

⟳



Sperm Processing Media3

Recommended application: Indirect Swim-up

To prepare the swim-up tubes transfer 4.5 ml medium into a new conical tube. Mix well with 1-3 ml liquefied semen.

Centrifuge the tube at 300-400 g for 10 minutes

1.

2. Aspirate the supernatant without dispersing the pellet and discard it.

Carefully overlay the pellet with equilibrated GM501 SpermActive or warmed GM501 SpermAir me-dium.

Put the tube in the CO2-incubator (GM501 SpermActive). Or heating cabinet (GM501 SpermAir) for 1 hour with not firmly closed top.

3.

4. Aspirate the supernatant contain-ing the motile sperms and fill it into a new conical centrifuge tube.

Resuspend the pellet with 1 ml equilibrated GM501 SpermActive or warmed GM501 SpermAir me-dium.


5.

6. Aspirate the supernatant without dispersing the pellet and discard it.

Resuspend the pellet with 1 ml equilibrated GM501 SpermActive or warmed GM501 SpermAir medium.


Repeat step 6.

Aspirate the supernatant and dis-card it.

For IVF resuspend the pellet in 0.1–1 ml equilibrated culture medium (e.g. GM501 Culture Media) to the desired sperm concentration, for ICSI resuspend it in GM501 Sperm-Active or GM501 SpermAir.Use this suspension for IVF/ICSI.

7.

8.



⟳

⟳

⟳



Sperm Processing Media

GM501 PVP

Product code: 1 x 1 ml 4 PVP 0001

5 x 0.2 ml 4 PVP 0001-5

GM501 PVP is a ready-to-use media to reduce the moti-lity of sperm making it easier to catch them with an ICSI pipette. It is possible to dilute the solution with HEPES-buffered handling media (e.g. GM501 Wash see pg. 1).


Composition:• GM501 PVP is 10 % dialysed Ph Eur grade polyvi-

nylpyrrolidone solved in an aqueous solution contain-ing physiologic salts, HEPES, Lactate, Pyruvate, Glu-cose and Human albumin solution.

Instructions for use:Standard procedure:• Warm the PVP solution to 37°C (Do not incubate in a

CO2-incubator, just warm it up to 37°C).• Place a small drop of PVP solution (5 µl - 10 µl) in a

dish and cover with GM501 MineralOil.• Add a small volume (1 µl - 2 µl) of washed sperm into

the centre of the PVP droplet.• Incubate for a few minutes to allow sperm to migrate

to the periphery of the droplet.• Select and recover the spermatozoa for injection.Alternativ procedure with extra washing step:• Warm the PVP solution and HEPES-buffered sperm

processing medium to 37°C. (Do not incubate in a CO2-incubator, just warm it up to 37°C.)

• Place a small drop of PVP solution (5 µl - 10 µl) and 1 or more small drops HEPES-buffered sperm proces-sing medium in a dish and cover with GM501 Mine-ralOil.

• Add a small volume (1 µl - 2 µl) of washed sperm into the centre of the PVP droplet.

• Incubate for a few minutes to allow sperm to migrate to the periphery of the droplet.

• Select the spermatozoa for injection and nick (break) the tail of the spermatozoon with the tip of the pipette.

• Transfer the spermatozoon into one of the HEPES-buffered sperm processing medium droplets and wash by transferring the sperm cell in and out of the sperm processing medium several times.

• Suck the sperm cell into the pipette and use for ICSI procedure.

Product specifications and quality control:• All raw materials are of highest available purity (European

Pharmacopoeia and/or USP standard), if applicable.

pH 7.20-7.90(Realease criteria: 7.20-7.60)




Viscosity at 25°C (cP) > 170

MEA (Blastocysts after 96 h in %)

≥ 80after 30 min of exposure


• The MSDS for GM501 PVP is available upon request and can also be downloaded from our website.

• GM501 PVP is manufactured according to the follow-ing specifications:

Sperm Processing Media 3





• GM501 PVP medium does not contain any antibiotics• The long term safety of ICSI, including the use of PVP,

on children born following this procedure is unknown.• Only for the intended use.






0344




GM501 Gradient

GM501 Gradient is a silane-silica based density gradient medium used in ART for separation and purification of highly motile human spermatozoa.


Instructions for use:• We advice to produce a 2-phase system from the

100 % gradient, comprising a 45 % and a 90 % grad-ient. You may prefer another mixing ratio which could be produced from the 100 % gradient as well (e.g. 45 %-70 %-90 %).

• Mix 9 parts of GM501 Gradient 100 % with 1 part of washing medium, to produce the 90% gradient.

• Mix 4.5 shares of GM501 Gradient 100 % with 5.5 sha-res of washing medium, to produce the 45 % grad-ient.

Note: Gradients should be prepared and repacked under sterile conditions (e.g. LAF-bench, ISO Class 5). For optimal results prepare the gradient media maximum 24 hours prior to use. Mix well after diluting the GM501 Gradient 100 %.



• A certificate of analysis is available for each batch

pH 7.20-7.90 (Release criteria: 7.20-7.60)

Osmolality (mOsm/kg)

300-330 (GM501 Gradient 100 %)310-340 (GM501 Gradient 45 %)320-350 (GM501 Gradient 90 %)

Density (g/ml) 1.1150-1.1250 (GM501 Gradient 100 %)1.1050-1.1150 (GM501 Gradient 90 %)

Viscosity (cP) < 1.65 (GM501 Gradient 90 %)< 1.75 (GM501 Gradient 100 %)


Endotoxin (EU/ml) < 0.5

Sperm Survival Test (%)

≥ 80 survival after 45 minutes exposure

of density selected sperm atozoa to the test medium

Mouse Embryo Assay (MEA)

not MEA tested


• The MSDS for GM501 Gradient is available upon re-quest and can also be downloaded from our website.

• GM501 Gradient is manufactured and tested accord-ing to the following specifications:

0482

Product code:GM501 Gradient 100 % (stock solution)1 x 50 ml 4 GM 501G-100-501 x 100 ml 4 GM 501G-100-1001 x 250 ml 4 GM 501G-100-2501 x 500 ml 4 GM 501G-100-500

GM501 Gradient 90 % (ready-to-use)1 x 10 ml 4 GM 501G-90-10 1 x 50 ml 4 GM 501G-90-50 1 x 100 ml 4 GM 501G-90-100 1 x 250 ml 4 GM 501G-90-250

GM501 Gradient 45 % (ready-to-use)1 x 10 ml 4 GM 501G-45-101 x 50 ml 4 GM 501G-45-501 x 100 ml 4 GM 501G-45-100 1 x 250 ml 4 GM 501G-45-250

Precautions and warnings:• All human, organic material should be considered po-

tentially infectious. Handle all specimens as if capable of transmitting HIV or hepatitis.

• Always wear protective clothing when handling speci-mens.

• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to a void possible contami-nation, even when GM501 Gradient 45 % and 90 % medium contain Gentamicin.





opening, when sterile conditions are maintained.• Content can not be re-sterilized after opening.• Do not use after expiry date.




1.

2.

3.

4.

5.

6.

7.

8.

9.

80-90 %

40-45 %

GM501 Gradient: Recommended application

Instruction for use with fresh semen samples

Preparation:We recommend one of the products from the GM501 MediaLine:• GM501 Gradient

Before use warm all components of the system and the samples to 37°C or to room temperature.For the better separation of the spermatozoa you should not li quefy the sample but increase the force of the centri-fuge, but not to more than 500 g.

⟳

⟳40-45 %

Pipet 2.5 ml of the lower density gradient (40-45 %) into a sterile disposable centrifuge tube.

Using a 3 cc syringe with a 21 g needle, layer 2.5 ml of the higher density gradient (80-90 %) under the lower density gradient (40-45 %) free of air bubbles.

Take care that the two layers are di-stinctly separated. This is done by placing the tip of the needle at the bottom of the centrifuge tube and slowly dispensing the higher den-sity gradient.

These two layers of density are sta-ble for about two hours.

Gently place 2.5 ml of liquefied se-men onto the upper layer using a transfer pipette or syringe.

Centrifuge at 350-400 g for 15-18 minutes.

In case, no pellet is visible after this step, centrifuge for another 3 mi-nutes.

Aspirate the supernatant.

Using a syringe, resuspend the pellet with 2-3 ml of fresh washing medium.

Centrifuge at 300 g for 8-10 minu-tes.

In case you want to gain higher sperm concentrations it is advisa-ble to centrifuge for the whole 10 minutes.

Aspirate the supernatant and re-peat the last two steps.

Finally remove the remaining liquid to have the pellet resuspended again in the desired amount for the use with the subsequent procedure of assisted reproductive medicine (e.g. IVF, ICSI, IUI).




Insemination Kit

The Insemination Kit is a complete system for simple and safe preparation and processing of human spermatozoa out of the ejaculate, for homologous and hetereologous intrauterine inseminations (IUI).

The Insemination Kit uses the self motility of the male germ cells to isolate motile spermatozoa in high concentrations. We recommend the use if the ejaculate is normozoospermic and slightly oligo- and/or asthenozoospermic.


One Kit comprises:• 1 x Vial with 2 ml of GM501 SpermAir Medium• 2 x 2 ml-Syringes (sterile)• 1 x Short cannula (sterile)• 2 x Long cannulas (sterile)• 1 x Ampulla rack• 1 x Insemination Catheter (sterile)• 1 x Instructions for use

Compostion of the GM501 SpermAir Medium:• NaCl, KCI, KH2PO4, MgSO4


D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-essential and essential Amino Acids, HEPES, Human serum albumin (5.00 g/liter), Gentamicin (10.00 mg/liter), Phenolred.

Product specifications and quality control of the GM501 SpermAir Medium:• All raw materials are of highest available purity (European

Pharmacopaia and/or USP standard), if applicable.• A certificate of analysis is available for each batch


• The MSDS for GM501 SpermAir is available upon re-quest and can also be downloaded from our website.

• GM501 SpermAir is manufactured according to the following specifications:

Product code: 1 x Kit4SA-KIT-002

1 x Kit 4SA-KIT-002 memo(Insemination Catheterwith memo effect)

Precautions and warnings:• GM501 SpermAir contains the antibiotic Genta micin

Sulfate. Appropriate precautions should be taken to en-sure that the patient is not sensitized to this antibiotic.

• Standard measures to prevent infections resulting from the use of medicinal products prepared from human blood or plasma include selection of donors, screening of individual donations and plasma pools for specific markers of infection and the inclusion of effective ma-nufacturing steps for the inactivation/removal of viru-ses. Despite this, when medicinal products prepared from human blood or plasma are administered, the possibility of transmitting infective agents cannot be totally excluded. This also applies to unknown or emer-ging viruses and other pathogens. There are no reports of proven virus transmissions with albumin manufactu-red to European Pharmacopoeia specifications by es-tablished processes.

• Therefore, handle all specimens as if capable of trans-mitting HIV or hepatitis. Always wear protective clothing when handling specimens.

• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contamination, even when GM501 SpermAir medium contains Genta-micin.





Disposal:• The products must be dispose like potential contami-

nated, specific to each country.

Storage instructions and stability:• Shelf life is 6 months from time of manufacture.• Store between 2-8°C. • Do not freeze before use.• Keep away from (sun) light.• The product can be used safely up to 7 days after ope-

ning, when sterile conditions are maintained.• Do not use after expiry date.

pH (at 37°C) 7.20-7.50




MEA (Blastocysts in %)

≥ 80(after 5 days of culture in the product covered by mineral oil, starting

from zygote stage)




1. Warm up the SpermAir vial to 37°C.

2. Remove metal cap from the stopper and desinfect the stopper´s surface with isopropylalcohol (70 %).

3. Insert the enclosed short cannula through the stop-per. It serves as pressure balance valve.

4. Aspirate liquefied, analyzed ejaculate into an enc-losed 2 ml-syringe and attach a long cannula.

5. Hold the syringe with its tip upwards to collect air in the upper part of the syringe and press it out.

6. Insert the syringe’s cannula (tip downwards) through the vial´s stopper until the tip attaches the bottom of the vial.

7. Now release the ejaculate slowly and carefully by pressing it out of the syringe and let it suspend un-der the preparation medium without mixing the two liquids.

8. After deflation remove the syringe with the cannula carefully while leading the tip along the inner wall of the vial. Discard the syringe and the cannula.

9. Now carefully place the vial´s neck into the rack´s fork and store the vial at 37°C in a incubator (no CO2)/ war-ming cabinet for least 45 minute and not longer than 3 hours.

10. After the time carefully take the vial out and turn it upright. Attach a fresh long cannula on the tip of a fresh 2 ml-syringe and insert it again through the des-infected stopper.

11. Aspirate 0.5 to 1.0 ml of the upper media layer and remove the syringe with the cannula. The syringe now contains the SpermAir-fraction with the isolated moti-le sperms. Until the insemination procedure place the syringe with the cannula with the attached protection cap of the cannula in a incubator (no CO2)/ warming cabinet by 37°C.

12. To inseminate remove the cannula from the syringe and attach the enclosed IUI-catheter at the tip of the syringe. Previous to the filling with the suspension 1 ml air is being aspirated into the syringe, so it is as-sured that the total probe will be spend during inse-mination into the cavum uteri. (Dead air volume of the catheter 1,3 mm3).

13. The position assistance is adjusted corresponding to the anatomical proportions determined before.

14. The catheter is inserted until the position assistance is being on the outer uterine orifice.

15. As soon as the requested position has been reached, the catheter will be turned, so that the marks on the grip are lying visible on top. In this way both of the lateral openings at the very end of the catheter are lined up towards the applicators orifice.

16. The suspension with the spermatozoa is injected slowly into the cave uteri.

17. Finally the catheter is slowly being extracted out of the uterus.

Advice:If the sperm is not liquefied sufficiently 30 minutes af-ter the ejaculation, liquefy it by aspirating it into a sterile disposable syringe (2 or 5 ml) and flushing it out serveral times.Before doing this, let disturbing rude particles sediment and do not aspirate them into the syringe.

It is recommended to analyse the sperm concentration prior to the insemination. For the insemination at least 2 million grade A spermatozoa should be present. An inse-mination with lower than 0,5 M/ml is not recommended. If performed optimally, the sperm suspension contains no or just very few immotile spermatozoa.

Insemination Kit: Recommended application



Cryopreservation MediaCryopreservation Media4

GM501 EmbryoStore

GM501 EmbryoStore is a set of ready-to-use media for freezing and thawing of human embryos between 2PN and 4 cell stage.


Composition:• GM501 EmbryoStore are propandiol based cryo-

preservation media which also contain PBS, sucose and human serum albumin (14-15 g/liter).

• GM501 EmbryoStore do not contain antibiotics.

Instructions for use:• The volumes of media that are necessary for freezing

or thawing must be mixed well and warmed up to room temperature before use.


ropean Pharmacopoeia and/or USP standard), if ap-plicable.


• The MSDS for GM501 EmbryoStore is available upon request and can also be downloaded from our web-site.

• GM501 EmbryoStore is manufactured and tested ac-cording to the following specifications:

pH 7.20-7.40



Product codes:1 x EmbryoStore Kit 4 EMF01_P_KIT11 x 10 ml EmbryoStore Freeze 4 EMF01_P_F1 x 10 ml EmbryoStore Thaw 1 4 EMF01_P_T11 x 10 ml EmbryoStore Thaw 2 4 EMF01_P_T21 x 10 ml EmbryoStore Thaw 3 4 EMF01_P_T3

Precautions and warnings:• Standard measures to prevent infections resulting from










0344


≥ 80

Osmolality (mOsm/kg)EmbryoStore Thaw 3

530-560



Cryopreservation Media 4Cryopreservation Media

GM501 EmbryoStore: Recommended application

Caution: because of density differences the em-bryos tend to float upwards and shrink like raisins.

Preparation:We recommend the following products from the GM501 MediaLine:• GM501 EmbryoStore Kit

Ensure all media are well mixed before use.

Freezing:1. Using a sterile pipette place 1 ml of EmbryoStore

Freeze medium in a centre well dish (at room tempe-rature).

2. Add the embryos (2PN stage) to the freezing medium and allow them to settle for about 30 seconds.

3. Load the embryos in straws leaving about 1/5 air in the straw.

4. Seal the straws and label with name, date and number of embryos.

5. Start freezing program within 5-10 minutes. Below is an example of a freezing protocol.

Temperature range

Freezing rate

Time

Phase 1 RT to +4°C -10°C/min. 2 min.

Phase 2 +4°C to -6°C -2°C/min. 5 min.

Phase 3 -6°C (autoseeding) 0°C/min. 10 min.

Phase 4 -6°C to -30°C -0,3°C/min. 80 min.

Phase 5 -30°C to-196°C -199°C/min. 1 min.

Thawing:1. Place in a 4-well culture dish 1 ml of each Embryo-

Store Thaw thawing solution (1, 2 & 3). This leaves 1 well empty to retrieve the frozen/thawed embryos.

2. Prepare a water bath at 37°C to thaw the straws. Pre-pare a 1 ml tuberculin-syringe by filling it with 0.8 ml of air first followed by 0.2 ml of EmbryoStore Thaw 1 medium.

3. Remove the straws from liquid nitrogen and leave at room temperature for about 5 seconds.

4. Submerge the straw in the water bath at 37°C for ano-ther 5 seconds (ensure no frozen part remains in the straw).

5. Empty the straw by opening both sides of the straw (above the empty well) and blowing the contents of the syringe through the straw.

6. Retrieve the embryos using a microscope and place them in EmbryoStore Thaw 1 thawing solution.

7. Transfer the embryos to EmbryoStore Thaw 2 thawing solution after 3-5 minutes.

8. After another 3-5 minutes the embryos are transferred to EmbryoStore Thaw 3 thawing solution. Leave for a further 3-5 minutes before proceeding.

9. At the final stage the embryos are transferred in IVF culture medium (e.g. GM501 Cult Media) for washing and further culture.




GM501 SpermStore

Product code: 1 x 20 ml 4 SCP-20

pH 7.20-7.90 (Release criteria. 7.20-7.60)

Endotoxin (EU/ml) < 0.25

Sterility sterile - SAL10-3

(Sterility Assurance Level)

Sperm Survival test (%) ≥ 80 survival after 45 minutes

exposure of untreated semen to the test medium

MEA not MEA tested

GM501 SpermStore is a medium for freezing human sperm including epididymal or testicular sperm. The intended users are IVF professionals (lab technicians, embryologists or medical doctors).

Composition:• GM501 SpermStore is a ready-to-use HEPES buffered

cryopreservation medium which also contains physi-ologic salts, glycine, dextrose monohydrate, lactate, glycerol, sucrose and human serum albumin (3.95 g/liter) to protect sperm from damage due to the freezing procedure.

Instructions for use:• Ensure all media are well mixed before use.• The volumes of media that are necessary for freezing

or thawing must be mixed well and warmed up to room temperature before use.

Before freezing:• In case of very low sperm concentrations it is advisable

to concentrate the sperm before freezing. This may in-crease sperm quality after thawing and will reduce the number of straws to be frozen.

Freezing:• Allow the semen to liquefy at room temperature for 30

minutes. Mix 1 ml of sperm with 0.7 ml SpermStore. Add the medium in drops while gently swirling. Cauti-on: to avoid cold-shock, make sure SpermStore is at room temperature.

• Leave the mixture for 10 minutes at room temperature for equilibration. Suck the sample/medium mixture into the freezing straws, leaving approximately 1.5 cm of air at the end of the straw. Seal the straws. Dry off indivi-dually with a linen free cloth.

• Shake to move the air-bubble to the centre of the straw.• Freeze vertically for 15 minutes, just above the level of

the liquid nitrogen. Store in liquid nitrogen. Thawing:• Remove as many straws as required from the liquid ni-

trogen. Place the samples in tap water for 5 minutes.• Cut off the end of the straw, place the open end inside

a container (e.g. a test tube) and tap the straw against the side of the container to allow complete evacuation of the mixture.

• Dilute the concentrated sperm in a suitable insemina-tion medium (at least 3 ml per 0.5 ml semen) and mix thoroughly. Centrifuge during 15 minutes at 300-350 g.

• Resuspend pellet in a suitable insemination medium. After thawing:• If necessary, use sperm prepation techniques after

thaw ing the semen to eliminate dead sperm cells and debris. Dilute the concentrated sperm in a suitable in-semination medium.

Product specifications and quality control: • All raw materials are of highest available purity (Europe-

an Pharmacopoeia and/or USP standard), if applicable.• A certificate of analysis is available for each batch upon

request from our website with respective lot number.• The MSDS for GM501 SpermStore is available upon

request and can also be downloaded from our website.• GM501 SpermStore is manufactured and tested ac-

cording to the following specifications:





• Only for the intended use.Pre-use checks:• Do not use the product if bottle, seal of the container





0344




GM501 VitriStore

GM501 VitriStore Freeze/VitriStore Thaw are a set of ready-to-use media for vitrification and thawing of human em-bryos.The intended users are IVF professionals (lab technicians, embryologists or medical doctors).

Composition:• GM501 VitriStore Freeze/VitriStore Thaw are DMSO/

ethylene glycol based vitrification media that also con-tain PBS, sucrose, Ficoll and human serum albumin (10-20 g/liter).

• GM501 VitriStore Freeze/VitriStore Thaw do not con-tain antibiotics.

Instructions for use:• Ensure all media are well mixed before use. We stron-

gly advice to read through all the steps of the vitrifica-tion/warming procedure before starting the procedure.




• The MSDS for the media are available upon request and can also be downloaded from our website.

• GM501 VitriStore Freeze/VitriStore Thaw are manufac-tured and tested according to the following specifica-tions:





• Only for the intended use.• The long term safety of embryo vitrification on child-

ren born following this procedure is unknown.





opening, when sterile conditions are maintained.• Do not use after expiration date.

References:• Stinshoff H., Wilkening S., Hanstedt A., Brüning K., Wrenzy-

cki C. (2011): Cryopreservation affects the quality of in vitro produced bovine embryos at the molecular level. Therioge-nology 76: 1433-1441

Product codes:1 x VitriStore Freeze Kit 4 VF_KIT 1 1 x 5 ml VitriStore Pre-vitrification Medium 1 x 1 ml VitriStore Freeze Medium 1 1 x 1 ml VitriStore Freeze Medium 2

1 x VitriStore Thaw Kit 4 VT_KIT 1 1 x 5 ml VitriStore Thaw Medium 1 1 x 1 ml VitriStore Thaw Medium 2 1 x 1 ml VitriStore Thaw Medium 3 1 x 1 ml VitriStore Thaw Medium 4

pH 7.20-7.40

Osmolality (mOsm/kg) 270-290 (VitriStore Freeze)

805-850 (VitriStore Thaw 1)



270-290 VitriStore Thaw 4)


≥ 80



0344



GM501 VitriStore: Recommended application (Freezing)

Preparation:Prepare the following products of GM501 VitriStore Freeze Kit:• 1 vial VitriStore Pre-vitrification Medium (PVM) (5 ml)• 1 vial VitriStore Freeze Medium 1 (VSF1) (1 ml)• 1 vial VitriStore Freeze Medium 2 (VSF2) (1 ml)

The media should be used in the order displayed above.

Ensure all media are well mixed before use. We strongly advice to read through all the steps of the vitrification/warming procedure before starting the procedure.

Preliminary steps:• In a 4-well dish fill the first well with 300 µl of Vit-

riStore Pre-vitrification Medium, the second with Vi-triStore Freeze 1 and the third with VitriStore Freeze 2 solution. Next open as many packs of vitrification devices as will be required for the vitrification step. Conveniently place the separate parts of the vitrifica-tion device on the workbench for easy access later in the procedure.

Freezing preparation:• Transfer the embryos from the blastocyst cell culture

medium into each of the VitriStore freeze solutions using the schemes.

Vitrification:1. Using an attenuated pipette or an equally suitable de-

vice, deposit maximum 2 blastocysts in a volume of approximately 0.3 µl of VitriStore Freeze 2 in the gut-ter of the tip of the vitrification straw.

2. Place the vitrification straw in the outer sheeth and seal it as indicated in the instructions for use of the vitrification device.

3. Plunge the sealed device into the liquid nitrogen.

*Before starting the vitrification procedure, in order to re-duce the negative effect of the blastocoel, expanded blas-tocysts should be collapsed by reducing artificially with a glass pipette the volume of the blastocoel.

Morulae/early blastocysts/PN-stages/early cleavage stages:

well 1 well 2 well 3

PVM VSF1 VSF2

2 min. 2 min. 30 sec.

Temperature of solutions and application at: Room temperature

Blastocysts/expanded blastocysts + artificial shrinking*:


PVM VSF1 VSF2


Temperature of solutions and application at: Room temperature

Blastocysts/expanded blastocysts:


PVM VSF1 VSF2


Temperature of solutions and application at: 37°C




Morulae/early blastocysts/PN-stages/early cleavage stages:


PVM VSF1 VSF2


GM501 VitriStore: Recommended application (Thawing)

Preparation:Prepare the following products of GM501 VitriStore Thaw Kit:• 1 vial VitriStore Thaw Medium 1 (5 ml) • 1 vial VitriStore Thaw Medium 2 (1 ml)• 1 vial VitriStore Thaw Medium 3 (1 ml)• 1 vial VitriStore Thaw Medium 4 (1 ml)

The media should be used in the order displayed above.

Ensure all media are well mixed before use. We strongly advice to read through all the steps of the vitrification/warming procedure before starting the procedure.

1. Remove the vitrifcation straw from the outer sheeth as indicated in the instructions for use of the vitrifica-tion device.

2. Immediately plunge the vitrification straw into pre-heated VitriStore Thaw Medium 1 (37°C) and leave in Thawing 1 medium for 3 minutes.

3. Transfer into VitriStore Thaw Medium 2 (37°C) and leave in this medium for 2 minutes.

4. Transfer into VitriStore Thaw Medium 3 (37°C) and leave in this medium for 2 minutes.

5. Finally transfer into VitriStore Thaw Medium 4 (37°C) and wash for at least 1 minute.

6. Transfer into blastocyst culture medium for continued cell culture (e.g. GM501 Cult Media).

Temperature of solutions and application at: 37°C

4-well-dish 4-well-dish

well 3 well 4

VitriStore Thaw Medium 3 VitriStore Thaw Medium 4

2 min 1 min

petri-dish 4-well-dish

well 1 well 2

VitriStore Thaw Medium 1 VitriStore Thaw Medium 2

3 min 2 min




Holding micropipettes are used for the fixation of oocytes, embryos or blastycysts and are therefore essential for all micromanipulation procedures in ART like ICSI, assisted hatching and polar body or blastomere biopsy.


Product specifications and quality control:• To meet international standards as well as the requi-

rements of the FDA, the Holding micropipettes are sterilised by gamma-radiation.

• The Holding micropipettes are prepared from borosi-licate glass tubing (O.D. 1.0 mm, I.D. 0.75 mm), total length 5.5 mm, polished opening, length arm 1 mm, bending angle 20°-40°. They may be ordered in 3 dif-ferent sizes small O.D. 80 µm, medium O.D. 100 µm and large O.D. 120 µm. Small and medium have po-lished opening an I.D. of 15-20 µm, large size has an I.D. 25 µm. The micropipettes are made straight or with bending angle.

• For better differentiation the Holding pipettes are de-livered in a green package.

• A mouse-embryo-test (MEA) is available for each batch upon request from our website with respective lot number.

Storage instructions and stability:• The Holding micropipettes are singly and sterile

packed and can be taken out easy, quick and safe of the protective package.

• The Holding micropipettes are sterilized for 3 years.

Holding micropipettes

small O.D.

Product code

code O.D.

µm

angle

degree

Polished opening

µm

Box

pcs.

001-80-20 001 80 20° 15 20

001-80-30 001 80 30° 15 20

001-80-35 001 80 35° 15 20

001-80-40 001 80 40° 15 20

001-80 001 80 straight 15 20

medium O.D.

Product code

code O.D.

µm

angle

degree

Polished opening

µm

Box

pcs.

001-100-20 001 100 20° 20 20

001-100-30 001 100 30° 20 20

001-100-35 001 100 35° 20 20

001-100-40 001 100 40° 20 20

001-100 001 100 straight 20 20

large O.D.

Product code

code O.D.

µm

angle

degree

Polished opening

µm

Box

pcs.

001-120-20 001 120 20° 25 20

001-120-30 001 120 30° 25 20

001-120-35 001 120 35° 25 20

001-120-40 001 120 40° 25 20

001-120 001 120 straight 25 20

Product code and technical specifications:

O.D. Polished opening

Customized production of pipettes available on request.

Micropipettes5

1023



medium O.D.

Product code

code O.D.

µm

angle

degree

Polished opening

µm

Box

pcs.

001-100-20 001 100 20° 20 20

001-100-30 001 100 30° 20 20

001-100-35 001 100 35° 20 20

001-100-40 001 100 40° 20 20

001-100 001 100 straight 20 20

large O.D.

Product code

code O.D.

µm

angle

degree

Polished opening

µm

Box

pcs.

001-120-20 001 120 20° 25 20

001-120-30 001 120 30° 25 20

001-120-35 001 120 35° 25 20

001-120-40 001 120 40° 25 20

001-120 001 120 straight 25 20

Product code and technical specifications: ICSI (intracytoplamic Sperm Injection) micropipettes are used to aspirate and inject the sperm directly into the oo-cyte.



rements of the FDA, the ICSI micropipettes are steri-lised by gamma-radiation.

• The ICSI micropipettes are prepared from borosilicate glass tubing (O.D. 1.0 mm, I.D. 0.78 mm), total length 5.5 mm, bending angle 20°-35° with lenght of arm 0.5 mm, beveled 35°, with I.D. of the tip 4.5-5.0 µm. The micropipettes are made with or without spike, strait or with bending angle and beveled 30°-40°.

• For better differentiation the ICSI micropipettes are delivered in a blue package.


Storage instructions and stability:• The ICSI micropipettes are singly and sterile packed

and can be taken out easy, quick and safe of the protective package.

• The ICSI micropipettes are sterilized for 3 years.

Injection micropipettes


ICSI micropipettes

Product code

code I.D.µm

angledegree

spike (s)

Boxpcs.

002-5-20-s 002 5 20° Yes 20

002-5-20 002 5 20° No 20

002-5-30-s 002 5 30° Yes 20

002-5-30 002 5 30° No 20

002-5-35-s 002 5 35° Yes 20

002-5-35 002 5 35° No 20

002-5-s 002 5 straight Yes 20

002-5 002 5 straight No 20

Large ICSI-Spermatid micropipettes are use for aspirati-on and injecting immature sperm directly into the oocyte. ICSI-Spermatid micropipettes has I.D. 7.0-8.0 µm, O.D. 9.0-10 µm of the tip.

ICSI-Spermatid micropipettes

Product code

code I.D.µm

angledegree

spike (s)

Boxpcs.

002-7-20-s 002 7 20° Yes 20

002-7-20 002 7 20° No 20

002-7-30-s 002 7 30° Yes 20

002-7-30 002 7 30° No 20

002-7-35-s 002 7 35° Yes 20

002-7-35 002 7 35° No 20

002-7-s 002 7 straight Yes 20

002-7 002 7 straight No 20with spike

I.D.

O.D.

withoutspike

O.D.

I.D.


Micropipettes 5Micropipettes

1023



Hatching micropipettes

Hatching micropipettes are used for the mechanical ope-ning of the Zona pellucida of embryos or blastocysts by Partial Zona Dissection (mechanical assisted hatching).



rements of the FDA, the Hatching micropipettes are sterilised by gamma-radiation.

• The Hatching micropipettes are prepared from boro-silicate glass tubing (O.D. 1.2 mm), total length 5.5 mm, bending angle 20°-45°, lenght of the arm 1.0 mm with short thin taper (S) or with long thin taper (L), sharp point. Hatching micropipettes may be ordered strait or with benging angle.

• For better differentiation the Hatching micropipettes are delivered in a pink package.


Storage instructions and stability:• The Hatching micropipettes are singly and sterile


• The Hatching micropipettes are sterilized for 3 years.


with short thin taper

Product code

code angledegree

taper Boxpcs.

003-20-S 003 20° short 20

003-30-S 003 30° short 20

003-35-S 003 35° short 20

003-S 003 straight short 20

with long thin taper

Product code

code angledegree

taper Boxpcs.

003-20-L 003 20° long 20

003-30-L 003 30° long 20

003-35-L 003 35° long 20

003-L 003 straight long 20

angle

1 mm

straight

60 µm

200 µm

40 µm

300 µm

Short thin taper (S)

Long thin taper (L)


Micropipettes5

1023




Biopsy micropipettes

Biopsy micropipettes are used to perform biopsies on the embryo (blastocyst) or the oocyte (polar body) for Preim-plantation Genetic Diagnosis - PGD.


Product specifications and quality control:• To meet international standards as well as the require-

ments of the FDA, the Biopsy micropipettes are steri-lised by gamma-radiation.

• The Biopsy micropipettes are prepared from borosi-licate glass tubing (O.D. 1.0 mm, I.D. 0.78 mm), total length 5.5 mm, bending angle 20°-45°, length of the arm 0.5 mm, with blunt opening (A) or beveled 40° and polished (B) and with I.D. 10, 15, 20, 30 and 35 µm. Biopsy micropipettes may be ordered strait or with benging angle.

• For better differentiation the Biopsy micropipettes are delivered in a yellow package.


Storage instructions and stability:• The Biopsy micropipettes are singly and sterile


• The Biopsy micropipettes are sterilized for 3 years.


Blunt opening A

Product code

code I.D.µm

angledegree

Blunt opening

Boxpcs

004-10-30-A 004 10 30° A 20

004-10-35-A 004 10 35° A 20

004-10-A 004 10 straight A 20

004-15-30-A 004 15 30° A 20

004-15-35-A 004 15 35° A 20

004-15-A 004 15 straight A 20

004-30-30-A 004 30 30° A 20

004-30-35-A 004 30 35° A 20

004-30-A 004 30 straight A 20

004-35-30-A 004 35 30° A 20

004-35-35-A 004 35 35° A 20

004-35-A 004 35 straight A 20

Blunt opening B

Product code

code I.D.µm

angledegree

Beveled Boxpcs.

004-10-30-B 004 10 30° B 20

004-10-35-B 004 10 35° B 20

004-10-B 004 10 straight B 20

004-15-30-B 004 15 30° B 20

004-15-35-B 004 15 35° B 20

004-15-B 004 15 straight B 20

004-30-30-B 004 30 30° B 20

004-30-35-B 004 30 35° B 20

004-30-B 004 30 straight B 20

004-35-30-B 004 35 30° B 20

004-35-35-B 004 35 35° B 20

004-35-B 004 35 straight B 20

I.D.

O.D.

A

I.D.

beveled 40°

O.D.

B


Micropipettes 5Micropipettes

1023



Denuding micropipettes

Denuding micropipettes are used for oocyte denudation or gamete and embryo handling.



rements of the FDA, the Denuding micropipettes are sterilised by gamma-radiation.

• The Denuding micropipettes are prepared from boro-silicate glass tubing (O.D. 1.2 mm, I.D. 0.75 mm), to-tal length 9.5 mm have a blunt opening and are sold straight.

• For better differentiation the Denuding micropipettes are delivered white (I.D. 120 and 150 µm), grey (I.D. 180 and 200 µm) and brown (I.D. 250 and 300 µm).


Storage instructions and stability:• The Denuding micropipettes are in a steril pack of

4 and can be taken out easy, quick and safe of the protective package.

• The Denuding micropipettes are sterilized for 3 years.


white

Product code

code I.D.µm

Box

005-120-C 005 120 40

005-150-C 005 150 40

grey

Product code

code I.D.µm

Box

005-180-B 005 180 40

005-200-B 005 200 40

brown

Product code

code I.D.µm

Box

005-250-A 005 250 40

005-300-A 005 300 40

150 µm 200 µm 300 µm

I.D. I.D. I.D.


8 Denudation-Tips6

1023



Product code and technical specifications: The DenuTips microtools for the manipulation and trans-fer of oocytes and embryos during IVF/ICSI procedures are made out of polycarbonate.


Product specifications and quality control:• The DenuTips are available in different sizes. • The DenuTips are flexible and unbreakable.• For better differentiation the Denu-Tips are available in

a variety of sizes each with a distinct colour.• A mouse-embryo-test (MEA) is available for each

batch upon request from our website with respective lot number.


Product code

size(µm)

colour Box

447125 125 blue 10

447135 135 yellow 10

447145 145 --- single

447155 155 black 10

447170 170 red 10

447200 200 purple 10

447275 275 green 10

Denu-Tips

The Gynettor is designed for comfortable and precise manipulation during denudation and transfer proce-dures. The vacuum system consists of silicone tube for continuous suck up and release of medium.

The intended users are IVF professionals (lab technici-ans, embryologists or medical doctors).

Instruction for use:• Carefully attach a sterile DenuTip to the opening

at the front of the device. • When the DenuTip fits please turn the front srew

clockwise to fix the DenuTip.

Storage instructions and stability:• The Denu-Tips are sterile and single packed.• The Denu-Tips are sterilized for 3 years.

Gynettor

Product specifications and quality control:• The holder is manufactured by mechanical machi-

ning from acetal copolymer resistant to hydrolysis, to strong alkali effects and to thermal oxidation degra-dation.

• The holder is manufactured of material harmless to health.

Storage instructions and stability:• All parts can be cleaned in the ultrasonic bath, retor-

ted or dehumidified up to the temperature of 100°C long-term and at 150°C short-term (1 hour).

• The Gynettor is reusable.

Product code: 1 x 006-H1

Denudation-Tips 6 Denudation-Tips



Laser Systems Micromanipulation Microscopes

Air Cleaning Spindle View Incubation

Work-Benches Laboratory control Safety

IVF Witness IVF Tracker Thax Findentity

Laboratory Equipment. Gynemed

May not be available in all German speaking countries!



Laboratory Equipment. Gynemed Miscellaneous 7

GM501 Flush

GM501 Flush is a ready-to-use medium for flushing the ovarian follicles during the aspiration of oocytes intended for extracorporeal fertilisation procedures. The medium is HEPES-buffered and contains Heparin to inhibit coagula-tion of blood. The medium does not contain human serum albumin (HSA).



.7H2O, NaHCO3, CaCl2.2

H2O, D(+)-Glucose anhydrous, Na-Lactate (50 % so-lution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-essential and essential Amino Acids, HEPES, Heparin.

Instructions for use:• Do not equilibrate GM501 Flush in a CO2-incubator,

just warm it up to 37°C. GM501 Flush is HEPES-buf-fered. Incubation in a CO2-incubator will lower the pH.

• Keep GM501 Flush medium at 37°C in a warming block throughout the entire process of oocyte reco-very and transport to the labor.

• Take about 20-50 ml medium at a time, using a rubber free syringe. Assemble needle and tubing set accor-ding to manufacturers instructions.

• Attach syringe containing GM501 Flush medium to the appropriate Luer, allowing connection to the lu-men of the needle.

• Aspirate the follicular fluid and flush the follicle with a volume of medium equivalent to the amount of follicu-lar fluid collected. Take care not to expand the follicle beyond its original size.

• Wash oocytes with GM501 Flush or pre-equilibrated culture medium, taking care to remove any blood clots and granulosa cells.

• Transfer oocytes to pre-equilibrated culture medium and store at 37°C in a 5-6 % CO2 environment.


ropean Pharmacopoeia and/or USP standard) if ap-plicable.

Product code: 1 x 50 ml 4 GM 501F-50

pH (at 37°C) 7.20-7.40





≥ 80


• The MSDS for the media are available upon request and can also be downloaded from our website.

• GM501 Flush is manufactured and tested according to the following specifications:

Precautions and warnings:• Always work under strict hygienic conditions (e.g.

LAF-bench, ISO Class 5) to avoid possible contami-nation.

• Always wear protective clothing when handling spe-cimens.










MiscellaneousMiscellaneous7

GM501 HSA

Product code: 1 x 5 ml 4 HSA-0005

GM501 HSA (10 % human serum albumin) is intended for use in assisted reproductive procedures which include ga-mete and embryo manipulation. These procedures include the use of GM501 HSA solution as a supplement for culture medium.


Composition:• 10 % (100 mg/ml) GM501 HSA in saline buffer.

Instructions for use:• To supplement the 50 ml culture medium (e.g. GM501

Basic or GM501 Basic with Gentamicin see pg. 9-10) with GM501 HSA to a protein end concentration of 1 %. Keep 45 ml culture medium in the bottle and add 5 ml of GM501 HSA. Mix well.

Product specifications and quality control:• GM501 HSA is obtained from blood of donors of certi-

fied blood banks according to international guidelines. The HSA is dialyzed and subsequently solved in saline buffer.

• The protein supplement is manufactured using the mo-dified Cohn-Oncley cold ethanol fractionation process. The final product is sterilized by filtration and additio-nally heated for 10-11 hours at 60°C.

• All raw materials are of highest available purity (Euro-pean Pharmacopoeia and/or USP standard), if appli-cable.


• The MSDS for GM501 HSA is available upon request and can also be downloaded from our website.

• GM501 HSA is manufactured and tested according to the following specifications:

pH (at 37°C, 6% CO2) 7.20-7.40





≥ 80














Miscellaneous 7Miscellaneous

GM501 Collagenase

Product code: 1 x 3 ml 4 COLL_AIR

GM501 Collagenase is a reagent for the digestion of hu-man testicular tissue obtained by testicular biopsy (TESE) for in-vitro examination procedures. Using Collagenase, it is possible to degrade testicular tissue in single cells to fa-cilitate the isolation of sperm cells.The intended users are IVF professionals (biologists, lab technicians, embryologists or medical doctors).

Composition:• Collagenase solved in HEPES-buffered medium.• NaCl, KCl, KH2PO4, MgSO4


D (+)-Glucose Anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-es-sential and essential Amino Acids, 0.5 % Human se-rum albumin, Gentamicin, Phenolred, HEPES.

• It has an activity of 1000 CDU/ml (Collagenase Diges-tive Units). The Collagenase is obtained from culture filtrates of clostridium histolyticum.

Instructions for use:• Do not equilibrate GM501 Collagenase in a CO2-incu-

bator, just warm it up to 37°C. GM501 Collagenase is HEPES-buffered. Incubation in a CO2-incubator will lo-wer the pH.




• The MSDS for GM501 Collagenase is available upon request and can also be downloaded from our website.

• GM501 Collagenase is manufactured according to the following specifications:

pH (at 37°C) 7.10-7.40



NOTE:• Performing LAL-endotoxin- and MEA-tests is not possi-

ble with these media as the activity of the Collagenase inactivates the enzymes (LAL) and damages the mouse embryos (MEA), which are used for these assays, res-pectively. The basic media is LAL- and MEA-tested.



• The GM501 Collagenase may appear cloudy if warmed up to 37°C. Such deterioration may occur occasionally without loss of efficiency and is not evidence of micro-bial contamination. It will disappear if GM501 Collage-nase is cooled down to room temperature.


• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contamination, even when GM501 Collagenase medium contains Gentamicin.




• Do not use the product if it becomes discoloured, clou-dy or shows any evidence of microbial contamination under cooled conditions.


opening, when sterile conditions are maintained.• Do not use after expiry date.Due to national legislation is this product not available in all countries!

References:1. Steiner S; Wöber M; Strohmer H; Plas E; Ebner T; Obruca A;

A new method to facilitate ICSI with testicular sperm: com-bining enzymatic digestion, accumulation and stimulation (in press)

Digestion of human testicular tissue obtained by testicular biopsy.

Preparation:We recommend the following products from the GM501 MediaLine:• GM501 Collagenase• HEPES-buffered sperm processing medium

Extract 1.5 ml GM501 Collagenase out of original bottle and fill into a 5 ml round-bottom tube.

Do not equilibrate GM501 Col-lagenase with lid open in a CO2-Incubator, just warm it up to 37°C. GM501 Collagenase is HEPES-buffered. Incubation in a CO2-In-cubator will lower the pH.

1.

For digestion of testicular tissue carefully pick the chosen tissue pieces with a fine syringe cannula. For easier handling, if necessary, fill the tissue suspension into a 60 mm petri dish. Let adhesive transport or cryo medium drop off as much as possible and transfer into Collage-nase tube.

Control by microscope whether vi-able semen cells are present. If not, digest further tissue sample for di-agnostic evaluation.

3.

4.

Close the tube completely. Now put the tube for 60 minutes in the incubator or ideally in a heat cabi-net for digestion of the tissue.

Suspend the digested tissue by ca-refully pipetting up and down. Un-der ideal conditions a suspension of single testicular tissue cells and free semen cells has been formed. If coarse tissue pieces are still visi-ble, repeat the process 2 for further 20-30 minutes.

5.

6.

Now centrifuge the tissue-cell-sus-pension and wash twice with 1–2 ml HEPES-buffered sperm proces-sing medium. Discard the obtained supernatants.

After the last centrifugation re-suspend the pellet in a small volume of 30–80 µl HEPES-buffered sperm processing medium. Add a few µl of this suspersion into the evaluation dish.

7.

8.

2.

If no motile sperms can be found Gynemed recommends the appli-cation of GM501 SpermMobil (pp. 41-42).

⟳

GM501 Collagenase: Recommended application

Miscellaneous7



Miscellaneous 7Miscellaneous



GM501 SpermMobil

Product code: 1 x 5 ml 4 GM 501SMOBIL5

1 x 1 ml 4 GM 501SMOBIL5-1

GM501 SpermMobil is a HEPES-buffered reagent contai-ning low bicarbonate. It is used for in-vitro examination of sperms of necrozoospermic ejaculates as well as of immotile sperms isolated from testicular tissue (TESE).

The intended users are IVF professionals (biologists, lab technicians, embryologists or medical doctors).



.2H2O, D(+)-Glucose Anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutami-ne, Water, non-essential and essential Amino Acids, Phenolred, HEPES, Theophylline.

Instruction for use:• Do not equilibrate GM501 SpermMobil in a CO2-in-

cubator, just warm it up to 37°C. GM501 SpermMobil is HEPES-buffered. Incubation in a CO2-incubator will lower the pH.




• The MSDS for GM501 SpermMobil is available upon request and can also be downloaded from our web-site.

• GM501 SpermMobil is manufactured according to the following specifications:

pH 7.20-7.60














References:1. Ebner T.D., Tews G., Mayer RB., Ziehr S., Arzt W., Costa-

moling W., Shebl O. (2011): Pharmacological stimulation of sperm motility in frozen and thawed testicular sperm using the dimethylxanthine theophylline. Fertil Steril 96: 1331-1336

Sperm Survival test (%) ≥ 80survival after 45 minutes exposure of density se-

lected spermatozoa to the test medium



Miscellaneous7

Activation of immotile sperms from necrozoospermic ejaculates as well as from testicular tissue (TESE = testicular-sperm extraction) by GM501 SpermMobil.

Preparation:We recommend one of the products from the GM501 MediaLine:• GM501 SpermMobil

Do not equilibrate GM501 SpermMobil in a CO2-incuba-tor, just warm up to 37°C.

To facilitate sperm-activation add 1.5-2.0 µl GM501 Sperm Mobil to the sperm-containing drop (ap-prox. 5.0 µl) in sperm preparation-medium inside the Petri-dish.

1.

GM501 SpermMobil should be added to the opposing side from where the sperms are to be aspirated.

3. Wait for 10 minutes. The activating effect initiates after a few minutes and lasts for one hour.The dish should be placed on a heating plate at 37°C during the diagnostic evaluation.

2.

GM501 SpermMobil: Recommended application

Miscellaneous 7



Miscellaneous

GM501 SpermFluid

Product code: 1 x 10 ml 4 GM 501FLUID10

GM501 SpermFluid is a HEPES-buffered reagent designed to dilute viscous semen samples for in-vitro procedures, examination and assessment of human sperm motility and concentration.


Composition:• Bromelaine solved in HEPES-buffered medium.• NaCl, KCl, KH2PO4, MgSO4


.2H2O, D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutami-ne, Water, non-essential and essential Amino Acids, 0.5 % Human serum albumin, Gentamicin, Phenolred, HEPES.

Instructions for use:• Do not equilibrate GM501 SpermFluid in a CO2-incu-

bator, just warm it up to 37°C. GM501 SpermFluid is HEPES-buffered. Incubation in a CO2-incubator will lower the pH.




• The MSDS for GM501 SpermFluid is available upon request and can also be downloaded from our web-site.

• GM501 SpermFluid is manufactured according to the following specifications:

pH (at 37°C) 7.20-7.50




the use of medicinal products prepared from human blood or plasma include selection of donors, scree-ning of individual donations and plasma pools for spe-cific markers of infection and the inclusion of effective manufacturing steps for the inactivation/removal of viruses. Despite this, when medicinal products pre-pared from human blood or plasma are administered, the possibility of transmitting infective agents cannot be totally excluded. This also applies to unknown or emerging viruses and other pathogens. There are no reports of proven virus transmissions with albumin manufactured to European Pharmacopoeia specifica-tions by established processes.


• Always work under strict hygienic conditions (e.g. LAF-bench, ISO Class 5) to avoid possible contamina-tion, even when GM501 SpermFluid medium contains Gentamicin.






opening, when sterile conditions are maintained.• Do not use after expiration date.


Reference:• Krebs T., Sollmann K., Maas D.H.A., Saymé N. (2012): Bro-

melase - A new way to reduce viscosity of human semen. 45th annual conference of Physiology and Reproduction and 37th Veterinary & Human Medicine conference in Berlin

• WHO laboratory manual for the Examination and processing of human semen (2010), 5th ed.: 14.



Dilute semen sample with GM501 SpermFluid in proportion 1:1 (for highly viscous, tenacious ejaculate we recommend to previously grind the ejaculate roughly!).

1.

Incubate the ejaculate for approx. 10 minutes at 37°C.

3.

4.

After 10 minutes you can use the liquefied semen sample for...

...diagnostic evaluation.

5. WHO laboratory manual-quote: These treatments may affect se-minal plasma biochemistry, sperm motility and sperm morphology, and their use must be recorded.Attention: To calculate the sperm concentration the dilution 1+1 (1:2) of semen with Bromelaine must be accounted.

2.

GM501 SpermFluid: Recommended application

Motility and concentration evaluation in viscous semen samples.

Preparation:We recommend the following products from the GM501 MediaLine:• GM501 SpermFluid

Do not equilibrate GM501 SpermFluid in a CO2-incubator, just warm up to 37°C.

Miscellaneous7 Miscellaneous

Miscellaneous



GM508 Cult-Active

Product code: 1 x 1 ml 4 GM 508CULT-active1

GM508 Cult-Active is a bicarbonate-buffer ed reagent de-signed to investigate oocytes of patients with failed ferti-lization after previous Intracytoplasmatic Sperm Injection cycles. GM508 Cult-Active is designed to investigate if fertilizati-on failure after previous ICSI-cycles is due to a deficient oocyte activation.


Composition• NaCl, KCl, KH2PO4, MgSO4


D(+)-Glucose anhydrous, Na-Lactate (50 % solution), Na-Pyruvate, EDTA, Alanyl-Glutamine, Water, non-es-sential und essential Amino Acids, DMSO, Ca2+-Iono-phore A23187.

Instructions for use• GM508 Cult-Active must be equilibrated 4 hours in

vial not firmly closed at 5-7 % CO2 and 37°C prior to use.




• The MSDS for GM508 Cult-Active is available upon request and can also be downloaded from our web-site.

• GM508 Cult-Active is manufactured and tested ac-cording to the following specifications:

pH (at 37°C, 6 % CO2) 7.00-7.60





≥ 80











References:1. Ebner T., Bulfon-Vogel S., Gruber I., Sonnenleitner U., Wö-

ber M. Staples P., Shebl O., Mayer R.B. Oppelt P. (2014): Successful treatment with Ca2+-ionophore in case of pre-vious developmental problems: a multicentre study. ESHRE P-148.

2. Ebner T.D., Montag M. (2011): Application of a ready-to-use ionophore increases rates of fertilization and pregnancy in severe male factor infertility. Fertil Steril 96: 34.

3. Montag M., Ebner T.D. (2011): Clinical application of artifi-cial oocyte activation: Result of a prospective multicenter study. Hum Reprod volume 26, Supplement 1 2011: Abs-tract Book: i106.

4. Montag M. et al. (2007): The relevance of oocyte activation in the era of ICSI. alpha no. 37

Miscellaneous 7



GM508 Cult-Active: Recommended application

Preparation:We recommend the following products from the GM501 MediaLine:• GM508 Cult-Active• GM501 Cult Media

Prepare a microdrop-dish in the following way:• Incubate the oocytes for 15 minutes in the pre-equili-

brated Ca2+-Ionophore GM508 Cult-Active drops.• Remove the oocytes from the GM508 Cult-Active

and wash them in culture media twice. This has to be done in a HEPES-or MOPS-free media, e.g. GM501 Cult media.

• Put the oocytes in your culture media for further cul-ture.

• Assess the development on specific moments in time.

Miscellaneous7

GM508 Cult-Active 30 µl/15 min

GM501 Cult Media

Miscellaneous

Gynemed Medizinprodukte GmbH & Co. KGLübecker Straße 9

23738 Lensahn

Germany

Tel.: +49 (0) 4363 90 32 90 Fax: +49 (0) 4363 90 32 9-19

[email protected]

Produkt_Broschüren: Broschüre_Fertility_Treatment: Gynemed_GM501_MediaLine_Katalog_EN_Rev10_00.indd/ Stand: 2015-01-26

This brochure is only a pre-publication and does not represent an agreement. All information in this brochure has been carefully prepared. Errors, misprints, Images and changes as well as variations may occur due to regulatory requirements.

Edition 02/2014 - MCH LAB• The MSDS for GM501 Wash is available upon request ... precautions...

Documents

Transcript of Edition 02/2014 - MCH LAB• The MSDS for GM501 Wash is available upon request ... precautions...