Course

1. Preparation and sterilization of MS medium
2. Surface sterilization and inoculation of explants (e.g., leaf, shoot tip)
3. Callus induction from explants
4. Micropropagation via shoot tip culture
5. Root induction in regenerated shoots
6. Preparation and sterilization of animal cell culture media
7. Thawing and revival of a mammalian cell line (e.g., HeLa or CHO)
8. Subculturing (passaging) of adherent cells
9. Trypan blue staining for viability and cell counting using a hemocytometer
10. Observation of cells under inverted microscope and cell counting

LEARNING OBJECTIVES

• Understand the fundamental principles and practices of plant and animal tissue culture.
• Gain hands-on experience with aseptic techniques and culture preparation.
• Learn standard procedures for initiating and maintaining plant and animal cell cultures.
• Develop skills in observing, analyzing, and interpreting cultured cell behavior under laboratory conditions.

COURSE OUTCOME

After completing the course, students shall be able to

CO1: Illustrate preparation and sterilization of culture media for plant and animal cell growth.

CO2: Perform surface sterilization and in vitro culture of plant explants for callus induction and regeneration.

CO3: Execute mammalian cell culture techniques such as revival, passaging, and viability assessment.

CO4: Perform quantification of cell growth and viability using hemocytometer and microscopy.

CO5: Demonstrate safe handling and maintenance of in vitro cultures, adhering to biosafety protocols.

Reference

1. Plant Tissue Culture: Theory and Practice“, Authors: S. S. Bhojwaniand M. K. Razdan
   Publisher: Elsevier
2. Introduction to Plant and Animal Tissue Culture“, Author: A. K. Bhattacharya
   Publisher: Kalyani Publishers