18BT44 Cell Culture Techniques syllabus for BT

CELL CULTURE LABORATORY DESIGN AND EQUIPMENTS:

Planning, construction and services; Layout; Sterile handling area; Incubation; Hot room; Air circulation; Service bench; Laminar flow; Sterilizer; Incubators; CO2 incubator; Culture Racks, Colony Counters, Refrigerators and freezers; Centrifuge; Inverted stage microscope; Magnetic stirrer; Liquid nitrogen freezers; Slow cooling system for cell freezing; Water bath; Autoclaves and hot air oven; Pipette washers; Water purification system; Fluid handling systems and other equipments; Washing, packing and sterilization of different materials used in plants, animals and microbial cell cultures; Aseptic concepts; Maintenance of sterility; Cell culture vessels

MEDIA AND REAGENTS :

Types of cell culture media for plants, animals and microbial cells; Ingredients of media; Physiochemical properties; Buffers; Oxygen; Osmolarity; Temperature; Balance salt solutions; Antibiotics, growth supplements; Conditioned media; Other cell culture reagents; Preparation and sterilization of cell culture media and other reagents.

ANIMAL CELL CULTURES TECHNIQUES:

History of animal cell culture; Different tissue culture techniques; Cell separation, disaggregation of the explants, mechanical and enzymatic disaggregation; Continuous cell lines; Organ culture, techniques, advantages, disadvantages, applications; Cell cultures, substrate culture and suspension culture; Primary cell culture; Secondary cell culture (cell lines); Development, characterization and maintenance of cell lines, Cryopreservation; Commercial scale production of animal cells; stem cells- fate mapping, application; Application of animal cell culture for in vitro testing of drugs;

PLANT CELL CULTURE TECHNIQUES:

Cellular Totipotency, And its Applications. Organogenesis, factors affecting organogenesis. Cytodifferentiation. Somatic Embryogenesis, Synthetic Seeds, Techniques for production of haploids, diploidization, production of double haploids and their Applications. Triploids production - Endosperm culture and Applications. Secondary metabolite production, selection of high yielding lines, elicitation, immobilization of cultures, hairy root culture and biotransformation. Factors affecting secondary metabolites, industrial application of secondary metabolites. Molecular farming.

MICROBIAL CELL CULTURE TECHNIQUES:

Auxotroph isolation - replica plating technique, Screening Preservation of microbial products. Production of antibiotics. Enumeration and screening of novel microbial secondary metabolites, strain improvement, Use of microbes in industrial waste treatment. Microbial leaching.

Course Outcomes:

At the end of the course the student will be able to:

• Differentiate between the various sources of cells to be used in cell culture techniques
• Correlate between different biological samples and understand the importance of different media in tissue culture
• Comprehend the applications of plant, animal and microbial cell culture in industry, healthcare and environment.

Question paper pattern:

• The question paper will have ten full questions carrying equal marks.
• Each full question will be for 20 marks.
• There will be two full questions (with a maximum of four sub- questions) from each module.
• Each full question will have sub- question covering all the topics under a module.
• The students will have to answer five full questions, selecting one full question from each module.

Textbook/s

1 Plant Cell Culture: A Practical Approach R.A. Dixon & Gonzales IRL Press 1994

2 Culture of animal cells-A manual of basic technique and specialized applications R. Ian Freshney Wiley Blackwell publishers 1983

3 Microbial Biotechnology Alexander N Glazer, Hiroshi Nikaido W H Freeman & Company 1995

Reference Books

1 Living resources for Biotechnology, Animal cells Doyle, R. Hay and B.E. Kirsop Cambridge University Press 1990

2 Plant Tissue Culture Sathyanarayana B N, IK Intl. Publishers 2007

3 Principle of Microbe & Cell Cultivation SJ Prit Blackwell Scientific co 1975