Long-term culture of mouse male germline stem cells under serum- or feeder-free conditions
Long-term culture of mouse male germline stem cells under serum- or feeder-free conditions
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DOI：
10.1095/biolreprod.104.036400
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发表时间：
2005-04-01
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;Miki, H;Shinohara, T
精原干细胞是人体中唯一能将遗传信息传递给下一代的干细胞。这些细胞可以在血清和饲养细胞的存在下长时间培养。然而，人们对调节精原干细胞自我更新分裂的因素知之甚少。在本研究中，我们探讨了建立缺乏血清或饲养细胞层的精原干细胞培养系统的可能性。精原干细胞可以在无血清条件下在小鼠胚胎成纤维细胞（mef）上扩增，也可以在无饲喂细胞的情况下在层粘连蛋白包被板上成功培养。然而，当血清和饲养细胞均缺失时，它们不能扩增。尽管在层粘连蛋白上培养的细胞与在饲养细胞上培养的细胞在表型上有所不同，但它们在至少6个月的时间里呈指数增长，并在移植到不育小鼠睾丸后产生正常、可育的后代。该培养系统将为了解精原干细胞的调控机制提供新的机会。
Spermatogonial stem cells are the only stem cells in the body that transmit genetic information to the next generation. These cells can be cultured for extended periods in the presence of serum and feeder cells. However, little is known about factors that regulate self-renewal division of spermatogonial stem cells. In this investigation we examined the possibility of establishing culture systems for spermatogonial stem cells that lack serum or a feeder cell layer. Spermatogonial stem cells could expand in serum-free conditions on mouse embryonic fibroblasts (MEFs), or were successfully cultivated without feeder cells on a laminin-coated plate. However, they could not expand when both serum and feeder cells were absent. Although the cells cultured on laminin differed phenotypically from those on feeder cells, they grew exponentially for at least 6 mo, and produced normal, fertile progeny following transplantation into infertile mouse testis. This culture system will provide a new opportunity for understanding the regulatory mechanism that governs spermatogonial stem cells.