An insight into the tissue An insight into the tissue culture technique and its culture technique and its

implicationimplicationDr.Samina Hyder HaqDr.Samina Hyder Haq

History of tissue cultureHistory of tissue culture 1885: Roux maintained embryonic chick cells in saline1885: Roux maintained embryonic chick cells in saline 1965:Ham introduced serum free culture medium that support 1965:Ham introduced serum free culture medium that support

living cellsliving cells 1965: Harris & Watkins successfully fused human and mouse 1965: Harris & Watkins successfully fused human and mouse

cells by viruscells by virus 1975: Kohler & Milstein produced the first Hybridomas capable 1975: Kohler & Milstein produced the first Hybridomas capable

of secreting monoclonal antibodies.of secreting monoclonal antibodies. 1978: Sato developed a serum free medium with a cocktail of 1978: Sato developed a serum free medium with a cocktail of

hormone and growth factorshormone and growth factors !982: human insulin is produced!982: human insulin is produced 1985: human growth factors were produced1985: human growth factors were produced 1986: lymphoblastoid gamaIFNlicences1986: lymphoblastoid gamaIFNlicences 1987:tissue type plasminogen activator(tPA) became 1987:tissue type plasminogen activator(tPA) became

commercially availablecommercially available 1989:Recombinant erythropoitin in trial1989:Recombinant erythropoitin in trial 1990:Recombinent products in trial (factorVI.HBsAg, Il2, EGF, 1990:Recombinent products in trial (factorVI.HBsAg, Il2, EGF,

mAbs)mAbs)

Plant Tissue culturePlant Tissue culture

MicropropagationMicropropagation

What is tissue cultureWhat is tissue culture

Organ culture Explant

Culture

Cell culture

Technique and instrument Technique and instrument

Carbon dioxide incubatorCarbon dioxide incubator

MicroscopeMicroscope

Tissue culture WareTissue culture Ware

Culture Media SterilizationCulture Media Sterilization

Cell countingCell counting

Changing MediumChanging Medium

Other miscellaneous EquipmentOther miscellaneous Equipment

Fridge Freezer for storing mediumFridge Freezer for storing medium Liquid nitrogen Container to freeze certain cell Liquid nitrogen Container to freeze certain cell

lineslines Incubator for warming up of the mediumIncubator for warming up of the medium Bench centrifuges to separate out cell pellet.Bench centrifuges to separate out cell pellet.

Types of cell cultureTypes of cell culture

1: Primary Cell culture1: Primary Cell culture 2: continuous Cell lines2: continuous Cell lines

Secondary or continuous Secondary or continuous Cell Lines Cell Lines

Attached Cell Lines    

Name Species and tissue of origin Morphology

MRC-5  (Prod. No. 84101801) Human lung Fibroblast

HELA  (Prod. No. 93021013) Human cervix Epithelial

VERO  (Prod. No. 84113001) African Green Monkey Kidney Epithelial

NIH 3T3  (Prod. No. 93061524) Mouse embryo Fibroblast

L929  (Prod. No. 85011425) Mouse connective tissue Fibroblast

CHO  (Prod. No. 85050302) Chinese Hamster Ovary Fibroblast

BHK-21  (Prod. No. 85011433) Syrian Hamster Kidney Fibroblast

HEK 293  (Prod. No. 85120602) Human Kidney Epithelial

HEPG2  (Prod. No. 85011430) Human Liver Epithelial

BAE-1  (Prod. No. 88031149) Bovine aorta Endothelial

SUSPENSION Cell linesSUSPENSION Cell linesSuspension Cell Lines    

Name Species and tissue of origin Morphology

NSO  (Prod. No. 85110503) Mouse myeloma Lymphoblastoid-like

U937  (Prod. No. 85011440) Human Hystiocytic Lymphoma Lymphoblastoid

Namalwa  (Prod. No. 87060801) Human Lymphoma Lymphoblastoid

HL60  (Prod. No. 98070106) Human Leukaemia Lymphoblastoid-like

WEHI 231  (Prod. No. 85022107) Mouse B-cell Lymphoma Lymphoblastoid

YAC 1  (Prod. No. 86022801) Mouse Lymphoma Lymphoblastoid

U 266B1  (Prod. No. 85051003) Human Myeloma Lymphoblastoid

SH-SY5Y  (Prod. No. 94030304) Human neuroblastoma Neuroblast

Storing and maintaining Continuous Storing and maintaining Continuous Cell LinesCell Lines

Advantages of Cell CultureAdvantages of Cell Culture

Cell types kept Constant and homogenous.Cell types kept Constant and homogenous. There is direct access to the cells so effect of There is direct access to the cells so effect of

toxicity of drug and chemicals studied without toxicity of drug and chemicals studied without being lost to other tissues and excreted.being lost to other tissues and excreted.

Replace/ reduce the number of animals.Legal, Replace/ reduce the number of animals.Legal, moral, ethycal isses?(animal rights Group).moral, ethycal isses?(animal rights Group).

Control of the environment (pH, osmotic Control of the environment (pH, osmotic pressure, temperature,oxygen and pressure, temperature,oxygen and Carbondioxide tension.) Carbondioxide tension.)

Disadvantages of Cell CultureDisadvantages of Cell Culture

Contamination can be Chemical Contamination can be Chemical (culture medium) Or Biological (culture medium) Or Biological (adding antibiotics).(adding antibiotics).

Finding a Happy Environment.Finding a Happy Environment. De-differentiationDe-differentiation Origin of CellsOrigin of Cells Major differences from Major differences from in-vivo(loss in-vivo(loss

of ECM)of ECM)

Implication for Cell CultureImplication for Cell Culture 1. Model system (many specialized functions are 1. Model system (many specialized functions are

restored)restored) 2.Toxicity Test (viability of cells)2.Toxicity Test (viability of cells) 3.Cancer research3.Cancer research VirologyVirology Cell Based manufacturingCell Based manufacturing Genetic councellingGenetic councelling Genetic engineringGenetic enginering Drug screening & developmentDrug screening & development Gene TherapyGene Therapy

Production of monoclonal antibodiesProduction of monoclonal antibodies

Tissue EngineeringTissue Engineering

Carbon nanotube Carbon nanotube scaffoldingscaffolding

Name of Name of scaffoldigscaffoldig

Made up ofMade up of

NanofiberNanofiber Like carbonLike carbon

TextileTextile PolyglycolidePolyglycolide

Gas FoamGas Foam Foam like Foam like structure due structure due to CO2 gasto CO2 gas

Growing cells in 3D Forming Growing cells in 3D Forming Tissue in BioreactorTissue in Bioreactor

Application of Monoclonnal Application of Monoclonnal AntibodiesAntibodies

IMMUNOLOCALIZATIONIMMUNOLOCALIZATION IMMUNOBlottingIMMUNOBlotting Cancer TreatmentCancer Treatment

ImmunolocalizationImmunolocalization

ImmunoblottingImmunoblotting

Cancer TreatmentCancer Treatment

Stem cell researchStem cell research

Embryonic Cell LinesEmbryonic Cell Lines

Adult Cell LineAdult Cell Line

Homeiopoitic Stem CellHomeiopoitic Stem Cell

Embryonic Cell linesEmbryonic Cell lines

Placental stem cellPlacental stem cell

Placental stem cells do not possess the antigenic Placental stem cells do not possess the antigenic properties, making rejection of the stem cells properties, making rejection of the stem cells impossible.impossible.

It has successfully used in treating various It has successfully used in treating various neurological disorders like Parkinson, neurological disorders like Parkinson, paralysis and stroke patients. paralysis and stroke patients.

Haematopoitic stem CellHaematopoitic stem Cell

Adult Mesenchymal Adult Mesenchymal Cell linesCell lines

Organ transplantOrgan transplant