Скачать или смотреть ANIMAL CELL CULTURE ||BIOTECHNOLOGY||CELL BIOLOGY||CELL CULTURE TECHNIQUES

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ANIMAL
CELL CULTURE
Cell in animal in an organised tissue matrix which require for their controlled growth and differentiation .
These cell from intact organism may be isolated or may be derived from a cell line or cell strain that has already been established, maintain and grown in vitro in culture media aseptically containing a suitable mixture of nutrient and growth factor. This process called Animal Cell Culture.
1907: ROSS HARRISON cultivate frog nerve cells in a lymph clot held by the ‘HANGING DROP’ method and observed the growth of nerve fibres in vitro for several weeks
 For sub culturing , the cells from primary cell culture are harvested/isolated and transferred to fresh culture media.
 Before sub culturing cells can be preserved in DMSO or glycerol by cryopreservation technique.
 Synthetic media prepared by adding several nutrient(organic and inorganic) , vitamins, salts, oxygen, co2 gas phase, serum protein, carbohydrates, co-factors etc.
 Serum containing media, serum free media, chemically defined media, protein free media etc.
 Serum free media
 Include serum extract , tissue extract or hydrolysate,growth factor, hormones, carrier protein like Albumin and Transferrin, lipids, metal ion , vitamins, polyamines etc
 Serum containing media
 Complete set of essential growth, hormones, attachment and spreading factor, binding and transporting protein human and bovine serum etc.
 It also suppress effect of shear stress to protect cells from physical damage .
 FCS, FBS, CS,HS, HoS
 1- plasma clots
 Prepared by mixing 15 drops of plasma with five drops of embryo extract in a watch glass .
 2- Biological fluids
 Amniotic fluid , aqueous humour from eye, pleural fluid.
 Serum is wildly used , obtain from adult human blood , placental cord blood, horse and calf blood
 3- tissue extract
 Chick embryo extract most commonly used, but bovine embryo extract , spleen , liver, bone marrow, leucocyte extract also used.
 PH – optimum ph between 7.2 to 7.4 is generally needed for mammalian cells. Phenol red is an internal indicator.
 Oxygen – cells depend upon glycolysis for the supply of o2.
 Temperature – optimal temperature 37
 Humidity- for cell growth 100/ humidity essential to reduce evaporation of the media.
 Antibiotic – Penicillin, Streptomycin , Gentamycin for Bacteria
 Nystatin – for fungi and yeast
 Laminar air flow hood
 Co2 incubator
 Centrifuge
 Refrigerator
 Cryo-strage container
 Liquid nitrogen container
 Haemocytometer
 water bath
 Flow cytometry
 Inverted microscope
 Dissecting microscope
 Glass wares and plastic wares


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