B.Sc.

Biotechnology Semester V BBT 513: Introduction to Animal Tissue Culture (ATC)

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45 L

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Introduction Introduction to animal tissue culture technique, historical review, definition of terms; tissue culture, cell culture, organ culture, primary cell culture, continuous cell lines; growth characteristics of cells growing in tissue culture; molecular basis of cell adhesion, anchorage dependent and independent cells. Laboratory requirements of ATC Infrastructure, equipments and materials; techniques for maintaining asepsis, different substrates used for in vitro growth of cells. Media Balanced salt solutions and growth media; physical and metabolic functions of different constituents of culture medium; role of carbon dioxide, serum, serum constituents and other supplements; commercially available important media. Basic techniques of mammalian cell culture Disaggregation of tissue and techniques for preparation of primary culture, routine subculturing and propagation of adherent cells and cells in suspension, development of cell line, introduction to cell banks. Cryopreservation Need for cryopreservation, stages of freezing of cells, revival. Quantitative methods Counting of cells using haemocytometer, electronic counting, methods for estimation of total DNA content, protein content, assays for measurement of cytotoxicity based on the rate of cell proliferation and metabolism. Organ culture Principles, methodology (dish, lens paper, sponge matrix, plasma clot and filter well inserts), advantages and limitations. Scalingup of animal cells in culture Scaleup of anchorageindependent and dependent cells, microcarriers, perfused monolayer cultures and bioreactors. Stem cell culture Sources, methodology and applications. Applications of animal cell culture Production of monoclonal antibodies and applications, recombinant proteins, growth factors, cell culture for vaccine production and drug toxicity. References 1. Animal Cell Culture Methods, Methods in Cell Biology, Vol. 57, Mather, J. P. and Bernes, D. Eds., Academic Press, New York (1998). 2. Animal Cell CulturePractical Approach, Masters, J. R.W. Ed., Oxford Press, London (2003). 3. Culture of Animal Cells, 5th Edn., Freshney, I., WileyLiss Publ., New Jersey (2006). 4. Principles and Practices of Animal Tissue Culture, Gangal, S., University Press (India) Pvt. Ltd., Hyderabad (2007).

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