Optimization of cryopreservation methods for somatic cells of the Tobet dog breed

Abstract

The Tobet dog breed, a national heritage of Kazakhstan, is threatened with extinction and conservation measures are urgently needed. This study presents a pioneering approach to preserve the genetic diversity and survival of this endangered breed through cryopreservation and cryobanking. We have optimised methods for cryopreservation of somatic cells (fibroblasts) isolated from skin explants of Tobet dogs and investigated the effects of in vitro cultivation on cell viability. Our results emphasise the importance of selecting appropriate culture media and cryoprotectants. Dulbecco's Modified Eagle Medium (DMEM) and ethylene glycol (EG) were found to be the most effective agents for increasing the growth rate and viability of fibroblasts after thawing. Through careful experimentation, including the evaluation of equilibrium and non-equilibrium cryopreservation techniques and the application of various cryoprotectants, we have established the first Tobet cryobank for somatic cells. The establishment of a cryobank represents a significant step towards the conservation of the Tobet dog breed and provides a model for the conservation of other endangered species. This study not only contributes to the preservation of Kazakhstan's cultural and biological heritage, but also opens up new avenues for the application of biotechnological approaches to wildlife conservation.