M.Sc. Biotechnology 2009-2010 M.

Sc Semester-III
MBT- 313: Animal tissue culture Total 45L 1 Introduction 5L Introduction to animal cell science and technology, brief review of the technology from its beginning to present date, definition of terms tissue culture, cell culture, organ, organotypic, histotypic cultures, primary, secondary and continuous cell lines, growth characteristic of cells growing in tissue culture, molecular basis of cell adhesion, anchorage dependent and independent cells. 2 Laboratory requirements of animal tissue culture technique 4L Requirements of infrastructure, equipment and material; techniques and quality control for maintaining asepsis, culture vessels and other substrates used for in vitro growth of cells Media 4L 3 Introduction to balanced salt solutions and growth medium, physical and metabolic functions of different constituents of culture medium, role of carbon dioxide, role of serum and supplements, commercially available important media formulations 4 Serum free defined media 2L Disadvantages of using serum in the medium, development of serumfree medium and their applications, commercially available serum free media 5 Basic techniques of mammalian cell culture 5L Disaggregation of tissue and techniques for preparation of primary culture, routine subculture and propagation, development of cell line, cryopreservation and revival, introduction to cell banks and its functioning 6 Measurement of viability and cytotoxicity 4L Estimation of cell viability, nature of assays for cytotoxicity testing, clonogenic assay, MTT based cell proliferation assays, anticancer drug screening Characterization of cultured cells 4L 7 Need for characterization, biochemical and molecular methods for characterization Scaling-up of animal cells in culture 4L Scaleup of anchorage independent and dependent cells, bioreactors, microcarriers, perfused monolayer cultures Cell cloning and transformation 4L Methods for cloning and transformation, tumerigenicity Organ culture 2L Principles and methodology of organ cultures, advantages and limitations of organ cultures, introduction to histotypic and organotypic cultures Applications of animal cell culture 7L Somatic cell fusion and production of monoclonal antibodies, cell culture products eg. viral vaccines, recombinant proteins, growth factors, stem cell

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cultures, embryonic stem cells and their applications References: 1. Reinert J.R. and Bajaj Y.P.S.(1997) Applied and fundamental aspects of plant cell, tissue and organ culture. Springer and Verlag, Berlin. 2. Bhojwani S. S. and Razdan M.K.( 2004) Plant Tissue Culture Theory and Practice, Elsevier publ., Amesterdam. 3. Adrian S., Nigel W. S. and Mark R. F. (2008) Plant Biotechnology: The Genetic Manipulation of Plants , 2nd eds., Oxford press. 4. Ian Freshney. Culture of Animal cells 5th eds.