History of animal tissue culture and natural surroundings for animal cell

History of animal tissue culture and natural

surroundings for animal cell

MADE BY : NEERAJ CHAUHAN

Introduction of ATC

• Animal Tissue Culture ?• Roux in 1885 for the first time maintained

embryonic chick cells in a cell culture• Cell culture was first successfully undertaken by

Ross Harrison in 1907.

Historical events in the development of cell culture

• 130-140 years old.• Arnold (1880) –showed that leucocytes can divide

outside body.• Roux (1885)- maintained embryonic chick cells in a

saline culture.• Jolly (1903)- studied behaviours of animal cells

immersed in serum lymph .• Ross Harrison (1907)- cultivated frog nerve cells in a

lymph clot and observed the growth of nerve fibers in vitro.

• Lewis (1911) - made the first liquid media consisted of sea water, serum, embryo extract, salts and peptones.

• Carrel (1913) - developed a method for maintaining cultures free from contamination.

• Rous and Jones (1916) – trypsinization and subculture of explants.

• Eagle (1955) – development of defined media.• Littlefield (1964) - introduced the HAT medium for

cell selection.• Ham (1965) - introduced the first serum-free medium

which was able to support the growth of some cells.

• Harris and Watkins (1965) - were able to fuse human and mouse cells by the use of a virus.

Factors which effect the choice choice of the substrate

1. Cell yield (cell production) –• For small scale production we use micro titration plates

multi well plates.

Micro titration plate

Multi well plates

• For large scale production we use flask and petri dishes

2.Whether the cells are monolayer/suspension culture -

• Monolayer culture Microtitration plate

• Suspension culture Flask

3.Venting – Airing to culture.

• Also called as aeration.

4.Sampling and Analysis – • Micro wells are used for sampling.• Two type of microscopes are used for

analysis.

Inverted microscopePhase contrast microscope

5.Uneven Growth - When r.p.m. is high during shaking than uneven growth comes.

6.Cost

1. pH- potential of H+ ion .

Optimum pH

Animal tissue – 7.4

Plant tissue – 5.5

Epidermal tissue – 5.5

Transformed tissue – 7-7.4

Fibroblast - 7.4- 7.7

2. Temperature – Optimum temperature

Animal – 37 F C

Birds – 38.5 F C

3. Gas Phase – Two phase CO2 - drops pH level • 5% required by the cells. O2 – 40-90% required• Some cells requires more O2 ,than extra O2

carrier sources added i.e Hb .

4.Osmolarity – Salt concentration of the cell

Animal - 290miliosmo /kg

Mice - 310milliosmo /kg

5.Foaming – Characteristic of suspension.

Drawbacks :- contamination occure.• Denaturation of protein.

• Interfare with the exchange of gas phase .

• To prevent foaming add antifoaming agent ex :- Pluronic F68,CMC( carboxy methyl cellulose)

6.Viscosity – Serum is added to increase viscosity.

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