IDENTIFICATION, SEPARATION AND ENRICHMENT OF SPERMATOGONIAL STEM CELLS FROM TIGER PUFFER (TAKIFUGU RUBRIPES)

This study investigated the isolation and purification of spermatogonial stem cells (SSCs) by single cell suspensions after enzyme digestion using testes of various months old tiger puffer. SSCs were enriched by Percoll discontinuous gradient. The SSCs were identified by optical observation and immunolocalization of Vasa protein. The majo-rity of SSCs at 14-month-old fish was type A spermatagonial cells that were significantly higher than those of the 22-month-old males (P<0.05). SSCs were mainly enriched in 10%—30% Percoll gradient belt, where the germ cells were mainly distributed at 14-month-old fish. These results indicate that 14-month-old male fish is a great source for SSCs using the current isolation method.