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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



Cultivation of Animal Cells En Masse in Bioreactor
The cells are cultivated in two different phases as freely suspended cells in liquid phase and immobilized cells on solid phase.

Suspension Culture. In suspen­sion culture the cells are dispersed in liquid medium and grow freely but not attached with any solid. The medium is agitated so that they should not form sediments. While adherent cells are permitted to get attached on solid sup­port and they grow as immobilized cul­ture. Therefore, it depends on choice of cells whether they are capable of growing freely in suspension or as immobilized culture (Anon, 1988).

A fully automated bioreactor is used that maintains the physicochemical and biological factors to optimum level and maintains freely the suspend­ed cells in an agitated low viscous liq­uid medium. The most suitable biore­actor used is a compact-loop bioreac­tor that consists of marine impellers (Fig. 6.6). It also relies on the inte­grated monitoring and control of phys­ical, chemical, biological and biochem­ical parameters.

As compared to bacteria, animal cells grow slowly. This results in bringing the cells in unfavourable metabolic state due to even small changes in culture. The main carbon and energy sources are glucose and glutamine. Lactate and ammonia are their metabolic products that affect growth and productivity of cells. Therefore, on-line monitoring of glucose, glutamate and ammonia can be carried out by on-line flow injection analysis (FIA) by using gas chromatography (GC), high performance liquid chromatography (HPLC), etc.

 
A compact loop bioreactor (diagrammatic).

Fig. 6.6. A compact loop bioreactor (diagrammatic).


Moreover, cell death also occurs due to certain reasons. Therefore, for measuring differences between viable and dead cells some enzyme activities are measured and correlated with cell density. Lactate dehydrogenase is used to distinguish cell density of viable cells (Fiechter, 1996).

 
     
 
 
     



     
 
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