A novel feeder-free culture system for expansion of mouse spermatogonial stem cells.
A novel feeder-free culture system for expansion of mouse spermatogonial stem cells.
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DOI：
10.14348/molcells.2014.0080
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发表时间：
2014-06
影响因子：
3.8
通讯作者：
Ko K
中科院分区：
文献类型：
--
作者：
Choi NY;Park YS;Ryu JS;Lee HJ;Araúzo-Bravo MJ;Ko K;Han DW;Schöler HR;Ko K
精原干细胞（SSC，也称为生殖干细胞）是自我更新的单能干细胞，可在睾丸中产生分化的生殖细胞。 SSC 可以从睾丸中分离出来，并在饲养细胞（通常是小鼠胚胎成纤维细胞）存在的情况下在体外长期培养。然而，SSC饲养培养系统的维护是繁琐的，因为每次传代培养时都需要准备饲养细胞。在本研究中，我们开发了一种基于基质胶的无饲养层培养系统，用于 SSC 的长期繁殖。尽管之前已经描述了几种没有饲养细胞的体外 SSC 培养系统，但我们基于基质胶的无饲养细胞培养系统具有时间和成本效益，并且保留了 SSC 的自我更新能力。此外，使用我们新开发的系统培养的 SSC 的生长速度与饲养层培养中的生长速度相当。我们证实，无饲养层培养的 SSC 在 mRNA 和蛋白质水平上均表达生殖细胞标志物。此外，无饲养层培养的SSCs的功能通过移植到生殖细胞耗尽的小鼠中得到证实。这些结果表明，我们新开发的无饲养层培养系统提供了一种在体外维持 SSC 和研究 SSC 的基础生物学（包括确定其命运）的简单方法。
Spermatogonial stem cells (SSCs, also called germline stem cells) are self-renewing unipotent stem cells that produce differentiating germ cells in the testis. SSCs can be isolated from the testis and cultured in vitro for long-term periods in the presence of feeder cells (often mouse embryonic fibroblasts). However, the maintenance of SSC feeder culture systems is tedious because preparation of feeder cells is needed at each subculture. In this study, we developed a Matrigel-based feeder-free culture system for long-term propagation of SSCs. Although several in vitro SSC culture systems without feeder cells have been previously described, our Matrigel-based feeder-free culture system is time- and cost- effective, and preserves self-renewability of SSCs. In addition, the growth rate of SSCs cultured using our newly developed system is equivalent to that in feeder cultures. We confirmed that the feeder-free cultured SSCs expressed germ cell markers both at the mRNA and protein levels. Furthermore, the functionality of feeder-free cultured SSCs was confirmed by their transplantation into germ cell-depleted mice. These results suggest that our newly developed feeder-free culture system provides a simple approach to maintaining SSCs in vitro and studying the basic biology of SSCs, including determination of their fate.
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DOI：
10.1098/rstb.2010.0026
发表时间：
2010-05-27
期刊：
Philosophical transactions of the Royal Society of London. Series B, Biological sciences
影响因子：
--
作者：
Phillips BT;Gassei K;Orwig KE
通讯作者：
Orwig KE
影响因子：
23.9
作者：
Ko, Kinarm;Tapia, Natalia;Schoeler, Hans R.
通讯作者：
Schoeler, Hans R.
DOI：
10.1073/pnas.0506970102
发表时间：
2005-10-04
影响因子：
11.1
作者：
Ryu, BY;Kubota, H;Brinster, RL
通讯作者：
Brinster, RL
影响因子：
14.8
作者：
Ko, Kinarm;Arauzo-Bravo, Marcos J.;Schoeler, Hans R.
通讯作者：
Schoeler, Hans R.
影响因子：
2.6
作者：
Nagano, M;Avarbock, MR;Brinster, RL
通讯作者：
Brinster, RL