Long-term culture of male germline stem cells from hamster testes
Long-term culture of male germline stem cells from hamster testes
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DOI：
10.1095/biolreprod.107.065615
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发表时间：
2008-04-01
影响因子：
3.6
通讯作者：
Shinohara, Takashi
中科院分区：
文献类型：
--
作者：
Kanatsu-Shinohara, Mito;Muneto, Tomomi;Shinohara, Takashi
精原干细胞是雄性动物精子发生的基础。我们最近成功地培养和基因工程小鼠精原干细胞，但很少有人知道在其他动物物种的精原干细胞的培养和生长的要求。本研究成功地从仓鼠睾丸中培养出精原干细胞。精原干细胞使用抗ITCA 6抗体纯化，并在胶质细胞系衍生的神经营养因子的存在下培养。细胞持续增殖至少1年。在此期间，他们使用慢病毒进行遗传修饰，并在移植到免疫缺陷裸鼠的睾丸后进行精子发生。然而，在替代异种受体中产生的生殖细胞没有分化超过精子细胞阶段，这些圆形精子细胞不能通过体外显微授精产生后代。这些结果表明，生殖细胞可能没有获得必要的生育能力在异种微环境的特性。然而，成功建立有利于仓鼠精原干细胞生长和维持的培养条件表明，这种技术可以扩展到其他动物物种，其中目前的遗传修饰技术是不可能的或效率低下的。
Spermatogonial stem cells provide the foundation for spermatogenesis in male animals. We recently succeeded in culturing and genetically engineering mouse spermatogonial stem cells, but little is known regarding the culture and growth requirements of spermatogonial stem cells in other animal species. In is study, we report the successful long-term culture of spermatogonial stem cells from hamster testes. Spermatogonial stem cells were purified using an anti-ITCA6 antibody and cultured in the presence of glial cell line-derived neurotrophic factor. The cells continued to proliferate for at least 1 year. During this period, they were genetically modified using a lentivirus and underwent spermatogenesis after transplantation into the testes of immunodeficient nude mice. However, germ cells generated in the surrogate xenogeneic recipients did not differentiate beyond the spermatid stage, and these round spermatids could not produce offspring through in vitro microinsemination. These results suggest that the germ cells may not have acquired characteristics necessary for fertility in the xenogeneic microenvironment. Nevertheless, the successful establishment of culture conditions conducive for hamster spermatogonial stem cell growth and maintenance indicates that this technique can be extended to other animal species in which current genetic modification techniques are impossible or inefficient.