History of animal tissue

culture and natural

surroundings for animal cell
 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 microscope
Phase 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 C
Birds 38.5 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.