Course

Practical

List of experiments1. Preparation and sterilization of MS medium2. Surface sterilization and inoculation of explants (e.g., leaf, shoot tip)3. Callus induction from explants4. Micropropagation via shoot tip culture5. Root induction in regenerated shoots6. Preparation and sterilization of animal cell culture media7. Thawing and revival of a mammalian cell line (e.g., HeLa or CHO)8. Subculturing (passaging) of adherent cells9. Trypan blue staining for viability and cell counting using a hemocytometer10. 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: Prepare and sterilize culture media for plant and animal cell growth using standard aseptic procedures.

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: Accurately observe and quantify cell growth and viability using a 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. Bhojwani and M.K. Razdan Publisher: Elsevier
2. Introduction to Plant and Animal Tissue Culture”Author: A. K. Bhattachary Publisher: Kalyani Publishers