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 and methods for evaluation of cell synchrony
8. New techniques in cell culture. Cell fusion.
9. Regulation of cell cycle
10. Cell signalling, growth factors, cell receptors, basic signalling pathways, transcription factors
11. basic mechanisms of tumorigenesis 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
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
to explain the regulating mechanisms in cell processes: regulation of cell cycle, signalling, senescence, apoptosis and connect them with the use of practical techniques in cell culture
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- Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P (2002):Molecular biology of the cell, Garland Publishing, New York.
Cooper G. M (2000): The cell: a molecular approach, ASM Press Washington DC, Sinauer Ass. Sunderland Massachusetts.
Ban J, Cerovec Ž (2004): Praktikum iz kulture animalnih stanica. Sveučilišni priručnik. Zagreb.
članci iz stručnih časopisa
- Masters J. R. W. (2000): Animal cell culture 3. iz. Oxford University Press, Oxford
Butler M (2004): Animal cell culture & technology, 2. iz. Bios Scientific Publishers, London i New York
Freshney R. I. (2000): Culture of animal cells: A manual of basic technique. 4. iz. Willey-Lys, J. Willey & Sons. New York
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