B. Sc.

in Medical Laboratory Technology

Practical Guide - Cell culture

Contents 1. 2. 3. 4. 5. 6. 7. 8. 9. Introduction to cell culture Equipment Cell culture media Serum Cell culture methods Preperation of primary cells Cryopreservation Quality control Health and safety

Objectives Student should be able to; 1. List types of cell lines and their properties. 2. Describe the applications of cell culture in virology and immunology. 3. Describe in brief the laboratory design and basic laboratory equipment required for cell culture. 4. Determine the cell viability and cell numbers in a cell suspension. 5. Demonstrate skills required for preparation, sterilization, storage and quality control of cell culture medium and reagents used in cell culture. 6. Describe preparation of glassware for cell culture. 7. Identify disposable plastic ware used commonly in cell culture. 8. Observe and describe the requirements and method of subculturing, maintenance and storage of cell lines. 9. Describe the common cell line contaminants and the precautions for elimination of contamination. 10. Describe how to handle cell line contamination. 11. Describe how to decontaminate spillages of cell culture material and disposal of cell culture wastes.

Cell lines and their properties 1. List types of cell cultures based on their cell division and briefly describe their properties. Primary cell cultures which are undifferentiated cells and closely resemble parent tissue. Secondary cell lines Continuous cell lines Semi continuous cell lines 2. Primary cell cultures are prepared from cells tissues or organs which are taken directly from an organism. Eg : kidney, chicken embryos, mouse embryos. List 2 methods of tissue disaggregation used in preparation of primary cell cultures. 1. Mechanical cutting with scissors, plunger, wire mesh 2. Treatment with enzymes such as trypsin and pronase. 3. List types of cell lines based on their adhesion properties Adherent cell lines Suspension cell lines 4. List 4 applications of cell culture in virology and immunology. Model system for - Isolation and diagnosis of pathogenic viruses. - Virus propagation - Study of cell physiology and biochemistry - Production of monoclonal antibodies. - Development and screening of antiviral drugs -large scale manufacturing of biological compounds (e.g., vaccines, therapeutic proteins).

5. What is a cell bank ? List 4 cell banks. The American Type culture collection European collection of animal cell cultures NIGMS human genetic mutant cell repository RIKEN

6. List 3 properties in cell lines that can undergo changes during prolong cell culture. 1. Change in morphology 2. Change in adherence 3. changes in the growth cycle 4. changes in expression of surface markers 5. changes in drug resistance

7. what is a cell strain ? A cell strain is a cell line selected by cloning from a parent cell line. The properties of the cell strain may be different from the parent cell line but all cells display same properties.

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9. Observe the design of the cell culture laboratory in the Department of Microbiology. Write down 5 important features of the laboratory design that you observe. Describe how these features are important. 1. 2. 3. 4. 5. Double doors Sink at the entrance Closed space Air conditioned. Equipped with all required equipment for cell culture and incubation so lowering the risk of contamination due to transport from one area to another. 6. Waste disposal