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

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...

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Detalhes bibliográficos
Autores: de Siqueira-Silva, Diógenes Henrique [UNESP], 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
Estado:Versão publicada
Data de publicação:2019
País:Brasil
Recursos:Universidade Estadual Paulista (UNESP)
Repositório:Repositório Institucional da UNESP
Idioma:inglês
OAI Identifier:oai:repositorio.unesp.br:11449/188991
Acesso em linha:http://dx.doi.org/10.1007/s10695-019-00631-8
http://hdl.handle.net/11449/188991
Access Level:Acceso aberto
Palavra-chave:Fish chimera
Germ cell technology
Spermatogonial proliferation
Surrogate production
Descrição
Resumo: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.