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Last update: RNDr. Tereza Tlapáková, Ph.D. (07.10.2020)
Practice takes place in person in shifts of 10 students. Theory and practice of cell and tissue culture. 1/3 of lectures, 2/3 practical class. Materials, sterilization, aseptic technique, media, supplements, disagregation, maintenance of lines, preservation, transformation, cell separation, measurement and quantitation, differentiation, culture of specific cells, gene transfer, karyology. |
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Last update: RNDr. Nataša Šebková, Ph.D. (21.02.2018)
Freshney, R. Ian: Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, 7th Edition. Wiley-Blackwell, 2016 (ISBN: 978-1-118-87365-6).
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Last update: RNDr. Tereza Tlapáková, Ph.D. (07.10.2020)
Credit from practical courses: 100% presence.
The exam requirements at the time of anti-epidemic measures remain (so far) the same.
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Last update: RNDr. Jana Rubešová, Ph.D. (14.09.2019)
Lectures:
1. Equipment of the laboratory: sterile area, laminar flow, incubators. Materials: glass, plastic, metals. Water: contamination, distillation, reverse osmosis, deionization, carbon filtration, sterilization.
2. Washing up. Sterilization. Aseptic techniques. Laboratory safety and biohazards. Contamination: mycoplasmas etc. The culture environment: osmolality, pH, buffering, temperature, gas phase.
3. Cell and tissue cultures: animal culture, feeder layer. Media and supplements: balanced salt solution, defined media, serum, serum-free media, selection of medium and serum, other supplements.
4. Disaggregation of the tissue and primary culture: isolation of the tissue, enzymatic disaggregation, collagenase, filtration, centrifugation. Maintenance of the culture: replacement of medium, cell lines, cloning, cell freezing and thawing.
5. Cell counting: hemocytometer, cell counter, calibration. Methods of cell separation: affinity methods, flow cytophotometry. Histochemistry, quantitation of DNA and proteins.
6. Karyology: chromosome preparation, chromosome banding and painting, hybridization.
7. Cell transformation: immortalization, contact inhibition, cell fusion, gene transfer.
Practical class: 1. Primary culture, preparation of medium. 2. Routine maintenance: replacement of medium, cell freezing and thawing. 3. Cell cloning in the presence of a feeder layer. 4. Chromosome preparation and banding. 5. Immunofluorescence.
The course is taught with the support of the project reg. number CZ.02.2.69/0.0/0.0/16_015/0002362 |