Preliminary study on testicular germ cell isolation and transplantation in an endangered endemic species Brycon orbignyanus (Characiformes: Characidae)

• Autor(a) principal: de Siqueira-Silva, Diógenes Henrique [UNESP]
• Data de Publicação: 2019
• Outros Autores: dos Santos Silva, Amanda Pereira [UNESP], da Silva Costa, Raphael [UNESP], Senhorini, José Augusto, Ninhaus-Silveira, Alexandre [UNESP], Veríssimo-Silveira, Rosicleire [UNESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNESP
• DOI: 10.1007/s10695-019-00631-8
• Texto Completo: http://dx.doi.org/10.1007/s10695-019-00631-8; http://hdl.handle.net/11449/188991
• topic: Fish chimera; Germ cell technology; Spermatogonial proliferation; Surrogate production

We aimed to develop a simplified protocol for transplantation of Brycon orbignyanus spermatogonial stem cells (SSCs) into Astyanax altiparanae testes. Brycon orbignyanus testes were enzymatically digested and SSC purified by a discontinuous density gradient. Endogenous spermatogenesis was suppressed in A. altiparanae using busulfan or by incubation at 35 °C water, and SSCs from B. orbignyanus labeled with PKH26 were injected into their testes via the urogenital papilla. Twenty-two hours post-transplantation, labeled spermatogonia were observed in A. altiparanae tubular lumen. After 7 days, spermatogonia proliferated in the epithelium, and 21 days post-transplantation, sperm was observed in the lumen. Of surviving host fish, nearly 67% of those treated with busulfan and 85% of those held in warm water showed labeled cells in host germinal epithelium. The present study standardized, by a simple and accessible method, germ cell transplantation between sexually mature Characiformes fish species. This is the first report of xenogenic SSC transplantation in this fish order.