1. An insight into the tissue culture technique and its implication Dr.Samina Hyder Haq

2. History of tissue culture 1885: Roux maintained embryonic chick cells in saline 1885: Roux maintained embryonic chick cells in saline 1965:Ham introduced serum free culture medium that support living cells 1965:Ham introduced serum free culture medium that support living cells 1965: Harris & Watkins successfully fused human and mouse cells by virus 1965: Harris & Watkins successfully fused human and mouse cells by virus 1975: Kohler & Milstein produced the first Hybridomas capable of secreting monoclonal antibodies. 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 1978: Sato developed a serum free medium with a cocktail of hormone and growth factors !982: human insulin is produced !982: human insulin is produced 1985: human growth factors were produced 1985: human growth factors were produced 1986: lymphoblastoid gamaIFNlicences 1986: lymphoblastoid gamaIFNlicences 1987:tissue type plasminogen activator(tPA) became commercially available 1987:tissue type plasminogen activator(tPA) became commercially available 1989:Recombinant erythropoitin in trial 1989:Recombinant erythropoitin in trial 1990:Recombinent products in trial (factorVI.HBsAg, Il2, EGF, mAbs) 1990:Recombinent products in trial (factorVI.HBsAg, Il2, EGF, mAbs)

3. Plant Tissue culture

4. Micropropagation

5. What is tissue culture Organ culture Explant Culture Cell culture

7. Technique and instrument

8. Carbon dioxide incubator

9. Microscope

10. Tissue culture Ware

11. Culture Media Sterilization

12. Cell counting

13. Changing Medium

14. Other miscellaneous Equipment Fridge Freezer for storing medium Fridge Freezer for storing medium Liquid nitrogen Container to freeze certain cell lines Liquid nitrogen Container to freeze certain cell lines Incubator for warming up of the medium Incubator for warming up of the medium Bench centrifuges to separate out cell pellet. Bench centrifuges to separate out cell pellet.

16. Types of cell culture 1: Primary Cell culture 1: Primary Cell culture 2: continuous Cell lines 2: continuous Cell lines

17. Secondary or continuous Cell Lines Attached Cell Lines NameSpecies and tissue of originMorphology MRC-5 (Prod. No. 84101801)Human lungFibroblast HELA (Prod. No. 93021013)Human cervixEpithelial VERO (Prod. No. 84113001)African Green Monkey KidneyEpithelial NIH 3T3 (Prod. No. 93061524)Mouse embryoFibroblast L929 (Prod. No. 85011425)Mouse connective tissueFibroblast CHO (Prod. No. 85050302)Chinese Hamster OvaryFibroblast BHK-21 (Prod. No. 85011433)Syrian Hamster KidneyFibroblast HEK 293 (Prod. No. 85120602)Human KidneyEpithelial HEPG2 (Prod. No. 85011430)Human LiverEpithelial BAE-1 (Prod. No. 88031149)Bovine aortaEndothelial

18. SUSPENSION Cell lines Suspension Cell Lines NameSpecies and tissue of originMorphology NSO (Prod. No. 85110503)Mouse myelomaLymphoblastoid-like U937 (Prod. No. 85011440)Human Hystiocytic LymphomaLymphoblastoid Namalwa (Prod. No. 87060801)Human LymphomaLymphoblastoid HL60 (Prod. No. 98070106)Human LeukaemiaLymphoblastoid-like WEHI 231 (Prod. No. 85022107)Mouse B-cell LymphomaLymphoblastoid YAC 1 (Prod. No. 86022801)Mouse LymphomaLymphoblastoid U 266B1 (Prod. No. 85051003)Human MyelomaLymphoblastoid SH-SY5Y (Prod. No. 94030304)Human neuroblastomaNeuroblast

19. Storing and maintaining Continuous Cell Lines

20. Advantages 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 toxicity of drug and chemicals studied without being lost to other tissues and excreted. 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). 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.) Control of the environment (pH, osmotic pressure, temperature,oxygen and Carbondioxide tension.)

21. Disadvantages of Cell Culture  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)

22. Implication for Cell Culture 1. Model system (many specialized functions are restored) 1. Model system (many specialized functions are restored) 2.Toxicity Test (viability of cells) 2.Toxicity Test (viability of cells) 3.Cancer research 3.Cancer research Virology Virology Cell Based manufacturing Cell Based manufacturing Genetic councelling Genetic councelling Genetic enginering Genetic enginering Drug screening & development Drug screening & development Gene Therapy Gene Therapy

24. Production of monoclonal antibodies

25. Tissue Engineering Carbon nanotube scaffolding Carbon nanotube scaffolding Name of scaffoldig Made up of Nanofiber Like carbon TextilePolyglycolide Gas Foam Foam like structure due to CO2 gas

26. Growing cells in 3D Forming Tissue in Bioreactor

27. Application of Monoclonnal Antibodies IMMUNOLOCALIZATION IMMUNOLOCALIZATION IMMUNOBlotting IMMUNOBlotting Cancer Treatment Cancer Treatment

28. Immunolocalization

29. Immunoblotting

30. Cancer Treatment

31. Stem cell research Embryonic Cell Lines Embryonic Cell Lines Adult Cell Line Adult Cell Line Homeiopoitic Stem Cell Homeiopoitic Stem Cell

32. Embryonic Cell lines

33. 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.

34. Haematopoitic stem Cell

35. Adult Mesenchymal Cell lines

37. Organ transplant