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TRAC 7: Animal and Human Cell Culture: Method and Applications
This lecture and laboratory course is structured to provide life scientists who are not experienced in cell culture with an introduction to principles and practices that will facilitate their ability to develop the use of in vitro systems. The course is predicated on the application of the most rigorous principles of quality control.
Lecture Topics: From Donor to Cell Lines: Comparison of Methods, Endothelial Cells from Aorta, Chick Embryo Culture, Human Skin Fibroblasts, Hepatic Cells; Explants and Cell Cultures; Contrasting Properties of Normal and Transformed Cells: Finite Life Span; Senescence and Telomeres; Transfection with Telomerase Gene; Mutagenesis and Carcinogenesis Assays; The in vitro environment: Sterility; Judicious use of Antibiotics, Mycoplasma Effects and Testing, The 2005 Consensus Document on Good Tissue Culture Practices; Low Density and Clonal Growth: Methods and use in Evaluation of Nutrients, Cell Proliferation, and Cytotoxicity; Cell Hybridization, Somatic Cell Genetics and Gene Mapping to Chromosomes; Cell Line Identification and Authentication: Karyology, Isoenzymes, Short Tandem Repeats. Cross-Contamination: Causes and Prevention; Karyotyping: Solid and G-banding. Chromosome Aberrations; Cell Culture and Biotechnology Interface: Transfection and Selection, Use of Genetically-Engineered Cells for Medical/Commercial Applications, Suspension and Mass Culture Techniques; Cell Culture Repositories: Starting and Maintaining a Cell Bank; Immortalized Cells.
Laboratory Topics: Subculturing 3 Different Cell Lines: Cell Counting and Cell Viability; Chick Embryo Culture; Gross Cytology of Cells: Staining, Morphological Changes During the Cell Cycle; Colchicine Synchronization; Metaphase Spreads; Solid Staining and G-banding; Cloning: Cloning by Limit Dilution and Plating efficiency; Transformed cells: growth patterns, growth in soft agar, and aggregate Formation; Chemoluminescent Assays: for Cell proliferation and Evaluation of Toxins; Transfection of Cells. Telomerase Assay; Apoptosis; Isoenzyme Assay for Detection of Interspecies Cross-Contamination
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