AT- 441: Animal  Tissue Culture                                                 (60) (Prerequisite: B Sc Life Science)   


Unit I                 (15) 

Animal cell culture: Introduction, applications and limitations. Laboratory organization: Design of ATC laboratory. Equipments used in animal tissue culture: Laminar Airflow Hoods, CO2 incubators, microscopes, refrigerators and deep freezers. Aseptic techniques in animal tissue culture. 


Unit II                 (15) 

Animal tissue culture media: Physical and nutritional requirements of cells, growth media and cell culture growth kinetics, different types of basal salt solution (BSS), minimal essential medium (MEM), antibiotic media, serum dependent defined media, serum independent defined media (for specific cell types), pH, bulk ions, trace ions,  CO2, O2 tension, ascorbic acid, vitamins and coenzymes. Types of contaminations in animal tissue culture. Protocols for routine characterization of cell lines. 


Unit III                          (15) 

Types of primary cells: epidermal, corneal, cervical, tumors cells, cerebellar, chondrocytes, osteoblasts, melanocytes and bone marrow cells. Basic techniques in animal tissue culture: Open and closed cell cultures, primary cell culture – isolation and separation of cells, viable cell count, maintenance of stock and regular cultures, antibiotic free stock culture. Types of animal cell cultures: monolayes, suspension, clonal culture, mass culture, micro carrier culture (monolayer), stem cell culture (ESC). 


Unit IV                (15) 

Cell fusion methods: Techniques involved in cell fusion, hybridoma cells: definition; preparation; properties and use of hybridoma technology, mini cells, micro cells and anucleated cells in fusion and their applications. Tissue engineering: capillary culture units; Techniques for culturing differentiated cells: reconstitution basement membrane rafts, feeder layers. Uses of animal cell culture technology: Mutant cell preparation; evaluation of chemical carcinogenicity and malignancy testing; metabolite production using animal cells and applications of pluripotent stem cells. 


Recommended Books: 1. Culture of Animal Cell: R. I. Freshney (Wiley-Liss) 2. Animal Cell Culture-Practical Approach: R. W. Jhon (Masters Oxford) 3. Biotechnology: U. Satyanarayana (Books & allied Pvt. Ltd.) 4. Methods in Cell Biology (Vol. 57)- Animal Cell Culture Methods: J. P. Mathon and D. Barnes (Eds) (Academic Press). 5. Mammalian Cell Biotechnology: A Practical Approach (1991): Butler, M. (IRL Press, Oxford)