This document discusses the history of animal tissue culture and factors that affect culturing animal cells. It notes that Roux in 1885 was the first to culture embryonic chick cells and Harrison in 1907 successfully cultured nerve cells. Key events over 130 years included development of defined media and serum-free media. Factors that impact cell culture choice include cell yield, whether cells are monolayer or suspension, venting, sampling needs, growth uniformity, and cost. Environmental factors like pH, temperature, gas phase, osmolarity, foaming, and viscosity must also be controlled to maintain optimal cell growth conditions.

1. History of animal tissue
culture and natural
surroundings for animal cell
MADE BY : NEERAJ CHAUHAN

2. 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.

4. 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.

5. • 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.

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

8. 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

9.  Multi well plates
• For large scale production we use flask and petri
dishes

10. 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.

11.  Inverted microscope
 Phase contrast microscope
5.Uneven Growth - When r.p.m. is high
during shaking than uneven growth comes.
6.Cost

13. 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

14. 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.

15. • 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.

16. Thanks