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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


Sterilization of Glassware, Equipments and Culture Media
The thoroughly washed glassware and equipments and carefully prepared culture media are sterilized by heat, steam or millipore filter paper (Table 6.4 ). The sterilized materials are removed when temperature cools down and used according to procedure adopted. For detailed description see In Vitro Culture Techniques: The Biotechnological Principles.


Table 6.4. Sterilization of glassware and equipments.

  Sterilization Items
  Autoclaving*
Apparatus containing glass and silicon tubing, disposable tips for micropipettes, dispensor tubing for pupet, glass bottles with screw caps, magnetic stirrer bars, screw caps, silicon tubing, topper-rubber, silicon, millipore filters, etc
  Dry heat** Glassware, glass coverslips, glass slides, instruments, Pasteur pipettes, pipettes, test tubes, etc.
*15 lb/in2 (=121°C) for 20 minutes; **160°C/ 1 hour.



Table 6.5. Sterilization of liquid by different methods.
  Sterilization Storage Solution
  Autoclave* Room temperature Agar, antibiotics, EDTA, glucose, glycerol, HEPES, lactalbumin hydrolysate, NaHCO3 solution, NaOH solution, phenol red, salt solution (with glucose), tryptose, water
  Autoclave 4°C Methocel
  Filter** Room temperature Glucose solution, HC1, NaHCO3 solution, NaOH solution,
  Filter -20°C Collagenase, antibodies, glutamine, sodium pyruvate solution, transferrin, trypsin, vitamines
  Filter-stacked filters 4°C Bovine serum albumin
  Filter-stacked filter -20°C Serum
  Steam (30 min at 100°C) 4°C Carboxylmethylcellulose
*15lb/in2 (=121°C) for 30 minutes, **0.2 mm pore size filter.

 
     
 
 
     



     
 
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