an insight into the tissue culture technique and its implication dr.samina hyder haq
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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