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Enrichment and In Vitro Culture of Spermatogonial Stem Cells from Pre-Pubertal Monkey Testes
Tissue Engineering and Regenerative Medicine ; (6): 557-566, 2017.
Artigo em Inglês | WPRIM | ID: wpr-646597
ABSTRACT
Spermatogonial stem cells (SSCs) are essential for spermatogenesis throughout the lifespan of the male. However, the rarity of SSCs has raised the need for an efficient selection method, but little is known about culture conditions that stimulate monkey SSC proliferation in vitro. In this study, we report the development of effective enrichment techniques and in vitro culturing of germ cells from pre-pubertal monkey testes. Testis cells were analyzed by fluorescence-activated cell sorting techniques and were transplanted into the testes of nude mice to characterize SSCs. Thy-1-positive cells showed a higher number of colonies than the unselected control after xenotransplantation. Extensive colonization of monkey cells in the mouse testes indicated the presence of highly enriched populations of SSCs in the Thy-1-positive sorted cells. Furthermore, monkey testis cells were enriched by differential plating using extracellular matrix, laminin, and gelatin, and then cultured under various conditions. Isolation of monkey testicular germ cells by differential plating increased germ cell purity by 2.7-fold, following the combinational isolation method using gelatin and laminin. These enriched germ cells actively proliferated under culture conditions involving StemPro medium supplemented with bFGF, GDNF, LIF, and EGF at 37 ℃. These results suggest that the enrichment and in vitro culture method proposed in the present study for harvesting a large number of functionally active monkey SSCs can be applied as the basis for efficient in vitro expansion of human SSCs.
Assuntos

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Espermatogênese / Células-Tronco / Testículo / Transplante Heterólogo / Técnicas In Vitro / Laminina / Haplorrinos / Colo / Fator de Crescimento Epidérmico / Fator Neurotrófico Derivado de Linhagem de Célula Glial Limite: Animais / Humanos / Masculino Idioma: Inglês Revista: Tissue Engineering and Regenerative Medicine Ano de publicação: 2017 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Espermatogênese / Células-Tronco / Testículo / Transplante Heterólogo / Técnicas In Vitro / Laminina / Haplorrinos / Colo / Fator de Crescimento Epidérmico / Fator Neurotrófico Derivado de Linhagem de Célula Glial Limite: Animais / Humanos / Masculino Idioma: Inglês Revista: Tissue Engineering and Regenerative Medicine Ano de publicação: 2017 Tipo de documento: Artigo