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  Section: General Biotechnology / Animal Biotechnology
 
 
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Animal Cell, Tissue and Organ Culture

 
     
 
Content
Requirements for animal cell, tissue and organ culture
  Substrates for cell culture
  Substrate treatment
  Culture media
    Natural media
    Synthetic media
  Sterilization of glassware, equipments and culture media
  Isolation of animal material (tissue)
    Disaggregation of tissue
    Establishment of cell culture
Cultivation of animal cell en masse in bioreactor
Immobilized cell culture
Insect cell culture
Somatic cell culture
Organ culture
  Organ culture on plasma clots
  Organ culture on agar
  Organ culture in liquid medium
  Whole embryo culture
Valuable products from cell cultures
  Monoclonal antibodies
  Production of commercial products from insect culture

Requirements for Animal Cell and Tissue Culture
Requirements for animal cell and tissue culture are the same as described for plant cell, tissue and organ culture (In Vitro Culture Techniques: The Biotechnological Principles). Desirable requirements are (i) air conditioning of a room, (ii) hot room with temperature recorder, (iii) microscope room for carrying out microscopic work where different types of microscopes should be installed, (iv) dark room, (v) service room, (vi) sterilization room for sterilization of glassware and culture media, and (vii) preparation room for media preparation, etc. In addition the storage areas should be such where following should be kept properly : (i) liquids-ambient (4-20°C), (ii) glassware-shelving, (iii) plastics-shelving, (iv) small items-drawers, (v) specialized equipments-cupboard, slow turnover, (vi) chemicals-sidled containers.

Substrates for Cell Growth
There are many types of vertebrate cells that require support for their growth in vitro otherwise they will not grow properly. Such cells are called anchorage-dependent cells. Therefore, a large number of substrates which may be adhesive (e.g. plastic, glass, palladium, metallic surfaces, etc.) or non-adhesive (e.g. agar, agarose, etc) types may be used as discussed below:

(i)

Plastic as a substrate. Disposable plastics are cheaper substrate as they are commonly made up of polystyrene. After use they should be thrown at proper place. Before use they are treated with gamma radiation or electric arc simply to develop charges on the surface of substrate. After cell growth its rate of proliferation should be measured. In addition, the other plastic materials used as substrate are teflon or polytetrafluoroethylene (PTFE), thermamox (TPX), polyvinylchloride (PVC), polycarbonate, etc. It should be noted that monolayer of cell must be grown. Moreover, plastic beads of polystyrene, sephadex and polyacrylamide are also available for cell growth in suspension culture.

(ii)

Glass as a substrate. Glass is an important substrate used in laboratory in several forms such as test tubes, slides, coverslips, pipettes, flasks, rods, bottles, Petti dishes, several apparatus, etc. These are sterilized by using chemicals, radiations, dry beat (in oven) and moist heat (in autoclave).

(iii)

Palladium as a substrate. For the first time palladium deposited on agarose was used as a substrate for growth of fibroblast and glia.

 
     
 
 
     



     
 
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