Course content
Course is giving theoretical and practical knowledge of the basic techniques in plant and animal cell culture.
Lectures
Animal cell culture: history of the cell culture and its application; basic composition of cell culture media and culture conditions. Primary cell culture. Methods of quantitation and characterisations of the cells. Growth curves and measurement of cell survival. Growth of tumour and normal cells and senescence. Methods of cell growth synchronisation. Cell fusion and differentiation in the culture. New techniques.
1. Introduction in animal cell culture: what is animal cell culture and where is it used today, comparison of cell culture with model organisms, history of cell culture,
2. conditions of cell culturing: types of cell culture medium and its components, types of substrates for cell growth, contamination, buffers used for pH conditioning, cryopreservation, types of cell lines, growth curves
3. Growth of cell colonies, methods of colony isolation, survival curves,
4. primary cell culture: preparation of primary culture, methods of separation of different cell types, evolution of the cell line
5. methods of cell quantitation: cell counting, determination of the cell viability by differential staining, flow cytometry.
6. Cell authentication and characterization: methods used for differentiating cells from diverse organisms, tissues and entities
7. Cell growth synchronisation: mitotic selection, synchronisation in G1 phase, inhibitors of S phase, double block in synchronization, methods for evaluation of cell synchrony
8. New techniques in cell culture. Cell fusion and differentiation. Basic characteristics of cancer cell growth in culture and senescence
Exercises
1. Introduction to cell culture laboratory and work in cell culture. Trypsinizing and counting cells. Seeding cells for growth curve and analysis of cell growth in culture by counting. Calculating the duplication time
2. Primary cell culture: preparation of fibroblast culture from mouse skin. Explantation technique and preparation of cell suspension by trypsinization
3. Influence of chemotherapeutics on the cell growth: growth curve obtained by counting the cells
4. Influence of chemotherapeutics on the cell growth: survival curve obtained by counting colonies
5. Cell synchronization: mitotic selection.
Outcomes
Animal cell culture
to analyse the growth conditions in the animal cell culture
to correlate the basic techniques and processes in the cell culture with the knowledge of cell biology
to apply the techniques of cell culture establishment and cell manipulation in the culture
to interpret and make conclusions about the data obtained by the experimental work during practical exercises
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- Ban J. Cerovec Ž. (2004): Praktikum iz kulture animalnih stanica. Sveučilišni priručnik. Zagreb.
Freshney R. I. (2000): Culture of Animal Cells: A Manual of Basic Technique. 4. iz, Willey-Lys, J. Willey & Sons. New York
Jelaska S. (1994): Kultura biljnih stanica itkiva. Školska knjiga, Zagreb.
- Masters J.R.W. (2000): Animal Cell Culture, 3rd iz. Oxford University Press, Oxford.
Butler M. (2004): Animal Cell Culture & Technology, 2nd iz, Bios Scientific Publishers, London i New York.
Alberts, B; Johnson, A; Lewis, J; Raff, M; Roberts, K; Walter, P. (2002) Molecular Biology of the Cell, 4th iz, Garland Science, New York i London.
Cooper, G. M. Hausman R. E. (2004) Stanica - molekularni pristup. Medicinska naklada Zagreb.
Lodish, H; Berk, A; Zipursky, S. L; Matsudaira, P; Baltimore, Darnell, J. E. (2000) Molecular Cell Biology. 4. izd. W. H. Freeman & Co, New York.
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