A Preliminary Technique for the Isolation and Culture of Brown Trout (Salmo trutta macrostigma, Dumeril, 1858) Spermatogonial Stem Cell

Abstract

This study was aimed to find a practical technique for isolation and culture spermatogonial stem cells from male brown trout (Salmo trutta macrostigma). Twelve wild juvenile male were obtained from Kılıç Trout Fish Farm (Kahramanmaraş, Turkey). The juveniles were taken alive to the aquaria unit and stored in a 1000-liter capacity fiberglass tank.  In order to identify the best size, age and testis structure of S.t. macrostigma for spermatogonial stem cell isolation and culture. Morphological and histological testis conditions were assessed. Fish were anesthetized with 0.04% 2-phenoxethanol. The surface of the fish was sterilized with 70% ethanol. Twelve fish were divided into two groups for enzyme digestion, and each group was divided into two replicates (three fish per replicate). Testis tissue of group one were digested by 0.25% trypsin- EDTA, and testis tissues of group two were digested by 0.05% trypsin-EDTA. At the end of the trial, first, the best age, size and weight of the male fish for spermatogonial stem cell isolation and culture were identified as 5+ month old, 12.13±1.5 cm, 19, 25±7.05 g respectively. Then, the highest spermatogonial stem cells were measured in the stage one and two of the testes. Finally, isolation and culture conditions were optimized for male S.t. macrostigma. Spermatogonial stem cell isolation and culture techniques were defined for fish in order to be used in surrogate reproduction technologies and gene transfer systems.