Anchorage-independent growth of mouse male germline stem cells in vitro
Anchorage-independent growth of mouse male germline stem cells in vitro
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DOI：
10.1095/biolreprod.105.046441
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发表时间：
2006-03-01
影响因子：
3.6
通讯作者：
Shinohara, T
中科院分区：
文献类型：
--
作者：
Kanatsu-Shinohara, M;Inoue, K;Shinohara, T
精子的发生起源于少量的精原干细胞，这些细胞位于基底膜上，在成年动物的整个生命周期中进行自我更新分裂，以支持精子的发生。尽管最近一项培养精原干细胞的技术的发展允许在体外进行自我更新分裂的繁殖，但关于精原干细胞是如何被调节的仍有很多未知。在这项研究中，我们发现精原干细胞可以以一种不依赖锚定的方式培养，这是来自其他类型自我更新组织的干细胞的特点。虽然培养的细胞生长缓慢（翻倍时间，约为4.7天），但它们表达了精原细胞标志物，并在至少5个月的时间内呈指数增长，达到1.5 × 10（10）倍的扩张。将培养的细胞移植到不育动物的精管中进行精子发生，并通过将培养细胞受体睾丸内发育的生殖细胞进行微授精获得可育后代。这些结果表明，精原干细胞可以进行不依赖于锚定的自我更新分裂，并表明干细胞在没有外源基质的情况下具有存活和增殖的共同特性。
Spermatogenesis originates from a small number of spermatogonial stem cells that reside on the basement membrane and undergo self-renewal division to support spermatogenesis throughout the life of adult animals. Although the recent development of a technique to culture spermatogonial stem cells allowed reproduction of self-renewal division in vitro, much remains unknown about how spermatogonial stem cells are regulated. In this study, we found that spermatogonial stem cells could be cultured in an anchorage-independent manner, which is characteristic of stem cells from other types of self-renewing tissues. Although the cultured cells grew slowly (doubling time, similar to 4.7 days), they expressed markers of spermatogonia, and grew exponentially for at least 5 months to achieve 1.5 x 10(10)-fold expansion. The cultured cells underwent spermatogenesis following transplantation into the seminiferous tubules of infertile animals and fertile offspring were obtained by microinsemination of germ cells that had developed within the testes of recipients of the cultured cells. These results indicate that spermatogonial stem cells can undergo anchorage-independent, self-renewal division, and suggest that stem cells have the common property to survive and proliferate in the absence of exogenous substrata.