Animal Cell Culture Research Training Module
Animal cell culture techniques is most widely used as model system to study basic cell biology and biochemistry, interactions between cells and disease forming agents and the effects of drugs. They have important application in generation of monoclonal antibodies and vaccine. It has been widely used for used for study in cancer biology, neurodegenerative diseases, cartilage regeneration, diabetes, virology, extraction of genetically engineered proteins, toxicity studies and drug screening and development. Most recently animal cell culture has found a major application in gene therapy. This is a basic training module intended for beginners to teach and train them in animal cell culture and equipment’s involved.
Objectives:
- It will be supported by theoretical classes and hands on practical classes which will include principle of the equipment’s and aseptic techniques essential for maintenance of animal cell culture. The participants will also have a chance to interact with experts and analyse their own samples (with prior approval).
Basic Techniques:
• Introduction to Good Laboratory Practices
• Media Preparation
• Cell Culturing
• Cell Counting
• Cryopreservation
• Cryo Revival
Screening and Evaluating Cell Viability, Cytotoxicity & Proliferation
• ALAMAR Blue assay
• MTT Assay
• SRB Assay
• Annexin V Assay
Cell based Staining and Imaging
• DAPI Staining
• Propidium Iodide staining
Cell based Molecular Expression and Analysis
• RNA Isolation
• cDNA Synthesis
• RT-PCR
Training Program Modules
Module-1
Introduction to the history and basics of Animal cell culture. Basics of Calculations; Good lab practices and aseptic techniques. Familiarization with the Animal cell culture laboratory and equipment’s and trouble shooting.
Module-2
Media and Reagent Preparation & Sterilization. Basics of cell culture initiation. Cell revival. Cell culturing. Cell counting. Cryopreservation. Basics of sub-culturing and growth curve. Cell line maintenance and characterization.
Module-3
Cell Counting. Cell Seeding for screening and Evaluating Cell Viability, Cytotoxicity & Proliferation.
Module-4
Processing of cells and preparations of chemicals for evaluation of cell viability, cytotoxicity & proliferation assay. Preparation of results and IC50 calculation. DAPI & PI staining.
Module-5
Preparation of Chemicals and processing of genomic DNA isolation from cancer cell lines. DNA Estimation; DNA agarose gel electrophoresis; Total RNA Extraction; cDNA synthesis from RNA
Module-6
Preparation of Chemicals and processing of Polymerase Chain Reaction from cDNA (Reverse Transcription-PCR); Agarose gel electrophoresis of PCR product; Total protein extraction.
Module-7
Preparation of Chemicals and processing of Agarose gel electrophoresis of PCR product; Total protein extraction; Protein estimation. SDS-PAGE electrophoresis of extracted proteins; Coomassie staining of protein gel.
Evaluation of trainees and certification
Evaluation of the training will be done and certificate will be issued to the successful candidates for the course..
