Transcript An insight into the tissue culture technique and its implication

An insight into the tissue
culture technique and its
implication
Dr.Samina Hyder Haq
History of tissue culture
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1885: Roux maintained embryonic chick cells in saline
1965:Ham introduced serum free culture medium that support
living cells
1965: Harris & Watkins successfully fused human and mouse
cells by virus
1975: Kohler & Milstein produced the first Hybridomas
capable of secreting monoclonal antibodies.
1978: Sato developed a serum free medium with a cocktail of
hormone and growth factors
!982: human insulin is produced
1985: human growth factors were produced
1986: lymphoblastoid gamaIFNlicences
1987:tissue type plasminogen activator(tPA) became
commercially available
1989:Recombinant erythropoitin in trial
1990:Recombinent products in trial (factorVI.HBsAg, Il2,
Plant Tissue culture
Micropropagation
What is tissue culture
Organ
culture
Explant
Culture
Cell
culture
Technique and instrument
Carbon dioxide incubator
Microscope
Tissue culture Ware
Culture Media Sterilization
Cell counting
Changing Medium
Other miscellaneous Equipment
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Fridge Freezer for storing medium
Liquid nitrogen Container to freeze certain cell
lines
Incubator for warming up of the medium
Bench centrifuges to separate out cell pellet.
Types of cell culture
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1: Primary Cell culture
2: continuous Cell lines
Secondary or continuous
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 lines
Suspension 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
Cell Lines
Advantages of Cell Culture
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Cell types kept Constant and homogenous.
There is direct access to the cells so effect of
toxicity of drug and chemicals studied without
being lost to other tissues and excreted.
Replace/ reduce the number of animals.Legal,
moral, ethycal isses?(animal rights Group).
Control of the environment (pH, osmotic
pressure, temperature,oxygen and
Carbondioxide tension.)
Disadvantages of Cell Culture
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Contamination can be Chemical
(culture medium) Or Biological
(adding antibiotics).
Finding a Happy Environment.
De-differentiation
Origin of Cells
Major differences from in-vivo(loss
of ECM)
Implication for Cell Culture
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1. Model system (many specialized functions are
restored)
2.Toxicity Test (viability of cells)
3.Cancer research
Virology
Cell Based manufacturing
Genetic councelling
Genetic enginering
Drug screening & development
Gene Therapy
Production of monoclonal antibodies
Tissue Engineering
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Carbon nanotube
scaffolding
Name of Made up of
scaffoldig
Nanofiber Like carbon
Textile
Polyglycolide
Gas Foam Foam like
structure due
to CO2 gas
Growing cells in 3D Forming
Tissue in Bioreactor
Application of Monoclonnal
Antibodies
IMMUNOLOCALIZATION
 IMMUNOBlotting
 Cancer Treatment
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Immunolocalization
Immunoblotting
Cancer Treatment
Stem cell research
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Embryonic Cell Lines
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Adult Cell Line
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Homeiopoitic Stem Cell
Embryonic Cell lines
Placental stem cell
Placental stem cells do not possess the antigenic
properties, making rejection of the stem cells
impossible.
It has successfully used in treating various
neurological disorders like Parkinson,
paralysis and stroke patients.
Haematopoitic stem Cell
Adult Mesenchymal
Cell lines
Organ transplant