Basics of Cell Culture细胞培养.ppt

* * * * Safety aspect in cell culture Possibly keep cultures free of antibiotics in order to be able to recognize the contamination Never use the same media bottle for different cell lines. If caps are dropped or bottles touched unconditionally touched, replace them with new ones Necks of glass bottles prefer heat at least for 60 secs at a temperature of 200 C Switch on the laminar flow cabinet 20 mts prior to start working Cell cultures which are frequently used should be subcultered stored as duplicate strains Other key facts…….? Use actively growing cells that are in their log phase of growth, which are 80-90% viable Keep exposure to trypsin at a minimum Handle the cells gently. Do not centrifuge cells at high speed or roughly re-suspend the cells Feeding sub culturing the cells at more frequent intervals then used with serum containing conditions may be necessary A lower concentration of 104cells/ml to initiate subculture of rapidly growing cells a higher concentration of 105cells/mlfor slowing growing cells Thanks * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Paras Yadav1, Annu Yadav1, P. Kumar1, J.S. Arora1, T.K.Datta1, S. De1, S.L. Goswami1, Mukesh Yadav2, Shalini Jain3, Ravinder Nagpal4 and Hariom Yadav3 1Department of Animal Biotechnology, 3Animal Biochemistry Division and 4Dairy Microbiology Division, National Dairy Research Institute, Karnal 132001 (Haryana), India; 2SOS in Chemistry, Jiwaji University, Gwalior-474011, M.P., India Basics of Cell Culture Introduction Cell culture is the process by which prokaryotic, eukaryotic or plant cells are grown under controlled conditions. But in practice it refers to the culturing of cells derived from animal cells. Cell culture was first successfully undertaken by Ross Harrison in 1907 Roux in 1885 for the first time maintained embryonic chick cells in a cell culture Historical events in the development of cell culture 1878: Claude Bernard proposed that physiological systems of an orga