Syllabus
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Course Code: IN-BTY -803 Course Name: Animal Cell Culture |
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| MODULE NO / UNIT | COURSE SYLLABUS CONTENTS OF MODULE | NOTES |
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| 1 | Biology of the Cultured Animal Cells, Tissue & Organ: Historical, Advantages and limitations - medical/pharmaceutical products of animal cell culture and their applications. Risks in a tissue culture laboratory and safety - biohazards. Facilities for animal cell culture: Infrastructure, Equipments including Biosafety Cabinets and Laminar Air Flow, Culture vessels – types (treated, Non-treated surfaces), the substrate, Nitrogen Container, CO2 incubator, Filters-sizes, types (for aqueous solution, for DMSO soluble solution). Biology and characterization of cultured cells-cell adhesion, proliferation, differentiation, morphology of cells and identification. Evolution of cell lines, development of continuous cell lines, dedifferentiation Culture Media: Balanced salt solutions, Complete media including proliferation, differentiation, Freezing and wash media. Brief discussion on the chemical, physical and metabolic functions of different constituents of culture medium. Role of CO2, Serum and Supplements. Serum free media and their application: advantages and disadvantages of serum and serum free media, replacement of serum and development of serum free media. Applications of Animal Cell Culture: Production of high value therapeutics (enzymes, hormones, monoclonal antibody, cytokines etc), virology, cancer research, gene therapy, drug development and cytotoxicity, animal cloning, genetic counseling, cryopreservation of cells. |
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| 2 | Primary Cell Cultures and Sub-cultures: types of primary cell culture, isolation of the tissue and preparation of primary cell culture, characteristics of limited life-span cultures, Techniques (mechanical disaggregation, enzymatic treatment, separation of viable and non-viable cells); Subculture and propagation, Criteria for subculture, Subculture of monolayer cells, growth cycle and split ratio, propagation and subculture in suspension ; Cell Lines and Characterization: Establishment and properties of continuous cell lines; Need for characterization, authentication (lineage or tissue markers), cell morphology, chromosome content, DNA content, RNA and protein expression, enzyme activity, antigen markers. Cell Cloning: Development of cloning techniques, dilution and suspension cloning, scaling up in suspension and monolayer, large scale production of cells using bioreactors, special requirement of cells growing at very low densities, uses of cloning; Organ culture - whole embryo culture Stem Cell Culture Embryonic and adult stem cells and their applications. Satellite Cells. Totipotent, Pluripotent and Multipotent stem cells. Induced Pluripotent Stem Cells (iPS) – Concept, Discovery, its impact on research in stem cell biology and in future therapy |
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