Syllabus
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
Annotation
During the anti-epidemic measures, the lectures take place online according to the schedule through the Microsoft Teams communication platform.
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.