L. A. Volkova
L.K. Ernst Federal Research Center for Animal Husbandry, pos. Dubrovitsy, 60, Podol’sk, Moskovskaya obl., 142132, Russian Federation
Abstract. Spermatogonia are classified as stem cells in the males’ testes, they are considered as a promising genetic material for storage in cryobanks in the framework of conservation and restoration the abundance of rare and endangered species, conservation and maintenance of the biodiversity in valuable species, breeds, lines and types of animals. The key aspect that determines the effectiveness of this technology is obtaining a pure population of individual types of spermatogenic cells. The aim of our research was to optimize the individual stages of isolation and maintenance of ram spermatogonia in culture. The objects of the study were the testes of the Romanov breed rams. The selection of this biomaterial was carried out by castration of males. Isolation of spermatogenic cells from ram testicles was carried out by successive mechanical and enzymatic treatments of testicular tissue using collagenase solution and trypsin solution. The optimal conditions for isolation and cultivation and the type of feeder layer were selected empirically. The best results were provided by the selection of biomaterial from males not older than 2 months. It has been shown that the high purity of the spermatogonia cell culture (up to 79%) is ensured by the preliminary removal other types of spermatogenic and somatic cells, including the largest population of Sertoli cells. Optimum properties for the cultivation of spermatogonia have a feeder layer formed by rams’ Sertoli cells. When using this type of feeder cells, the attachment of spermatogonia to the feeder layer was observed on the 1st-2nd days of cultivation, followed by the formation of colonies by the 6th day of cultivation.
Key words: Ovis, ram, cell culture, spermatogenic cells, spermatogonia.
Author Details: L.A. Volkova, Cand.Sc. (Biol.), senior research fellow (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.).
For citation: Volkova LA. [Isolation and characterization of ram spermatogonia] Dostizheniya nauki i tekhniki APK. 2022;36(7):67-70. Russian. doi: 10.53859/02352451_2022_36_7_67.