Cryopreservation and transplantation of germ cells in fish provides a suitable tool for preserving genetic information. By method of surrogate reproduction, the offspring with characters of the chosen donor can be obtained. In this case of our commercially important species common carp. However, for the successful cryopreservation of the germ cells, a suitable protocol for each species must be established. Several cryoprotectants were tested. The best of them, Me2SO, regarding the viability of spermatogonia, was tested for its different concentrations depending also on the rate of freezing. Further testing, related to the effect of tissue size, incubation time and added sugar, was performed. The result of the assay identified best cryomedium composed of 2.5M dimethylsulfoxide, added sugar of 0.3M glucose, 1.5% BSA and 25nM Hepes dissolved in PBS. The most suitable size of tissue was 100 mg, incubation time was 30 min and coolig rate was -1 ° C/min. This protocol ensures the highest viability rate of cryopreserved spermatogonia of common carp. The second part of the work was to verify the success of the transplantation of cryopreserved and fresh spermatogonia into a suitably chosen recipient, the goldfish, which shares similar reproductive characteristics with carp, but also offers reduction of space requirements or resistance to koi-herpes virus. The transplanted germ cells colonized the germ line and started gametogenesis in 42.5% (cryopreserved spermatogonia) and 52.5% (fresh spermatogonia) goldfish recipients, which demonstrated that the transplantation of cryopreserved spermatogonia of common carp can be successfully achieved.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:375951 |
| Date | January 2018 |
| Creators | FUČÍKOVÁ, Michaela |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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