Sendai virus-mediated transduction of mammalian spermatogonial stem cells.
Sendai virus-mediated transduction of mammalian spermatogonial stem cells.
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仙台病毒介导的哺乳动物精原干细胞转导。
DOI：
10.1093/biolre/ioy192
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Shinohara T.
中科院分区：
文献类型：
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作者：
Watanabe S.;Kanatsu-Shinohara;M.;Shinohara T.
精原干细胞（SSCs）是精子发生的基础。然而，由于其数量少且自我更新缓慢，转染ssc的成功率有限。虽然几种病毒载体可以感染SSCs，但基因组整合和无法维持长期基因表达阻碍了SSCs的研究。本文报道仙台病毒（SV）成功感染SSC，这是副粘病毒科的一种RNA病毒。SV有效地转导了种系干（GS）细胞，培养了具有丰富SSC活性的精原细胞，并维持了至少5个月的基因表达。它还感染从成年睾丸中新分离的ssc。在精子移植到不育小鼠的精管后，转染的GS细胞重新启动了精子发生，这表明SV转染不会干扰精子发生的进程。另一方面，将SV微注射到未成熟小鼠的精管中，可转导ssc和Sertoli细胞，但间质病毒注射不能转导间质细胞或小管周围细胞。异种精原细胞移植后鉴定出SV感染的仓鼠GS细胞和新鲜分离的兔或猴ssc样细胞，表明SV可以转导几种哺乳动物的ssc。因此，SV是一种有用的载体，可以转导ssc和Sertoli细胞，并克服与其他病毒载体相关的问题。
Spermatogonial stem cells (SSCs) provide the foundation of spermatogenesis. However, because of their small number and slow self-renewal, transfection of SSCs has met with limited success. Although several viral vectors can infect SSCs, genome integration and an inability to maintain long-term gene expression have hampered studies on SSCs. Here we report successful SSC infection by Sendai virus (SV), an RNA virus in the Paramyxoviridae. The SV efficiently transduced germline stem (GS) cells, cultured spermatogonia with enriched SSC activity, and maintained gene expression for at least 5 months. It also infected freshly isolated SSCs from adult testes. The transfected GS cells reinitiated spermatogenesis following spermatogonial transplantation into seminiferous tubules of infertile mice, suggesting that SV transfection does not interfere with spermatogenesis progression. On the other hand, microinjection of SV into the seminiferous tubules of immature mice transduced SSCs and Sertoli cells, but did not transduce Leydig or peritubular cells by interstitial virus injection. SV-infected hamster GS cells, and freshly isolated rabbit or monkey SSC-like cells were identified following xenogeneic spermatogonial transplantation, suggesting that SV transduces SSCs from several mammalian species. Thus, SV is a useful vector that can transduce both SSCs and Sertoli cells and overcome problems associated with other viral vectors.