Enrichment of Mouse Spermatogonial Stem Cells by the Stem Cell Dye CDy1.
Enrichment of Mouse Spermatogonial Stem Cells by the Stem Cell Dye CDy1.
复制标题
通过干细胞染料 CDy1 富集小鼠精原干细胞。
DOI：
10.1095/biolreprod.115.135707
复制
发表时间：
2015
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;Morimoto H;Shinohara T.
精原干细胞（ssc）由一小群具有自我更新能力的生殖细胞组成。先前的研究表明，ssc与其他自我更新组织中的干细胞具有一些共同特征，包括表面标记和增殖机制。然而，由于ssc数量少，缺乏ssc特异性标记物，严重阻碍了ssc的研究。在本研究中，我们研究了CDy1和Rh123的效用，它们都用于收集几种自我更新组织中的干细胞。CDy1染色的种系干（GS）细胞，体外培养的精原细胞富含SSC活性，无毒性作用。与先前报道的干细胞特异性染料不同，CDy1在GS细胞培养和成熟成年睾丸中也可用于SSCs的富集。经基于cdh1的磁性细胞选择和CDy1染色后，精原细胞移植显示66.7个细胞中的~ 1具有SSC活性。相比之下，尽管Rh123先前成功地从隐睾收集了ssc，但不可能从GS细胞培养和野生型睾丸中恢复ssc。因此，CDy1染色将为SSCs的富集提供一种有用的策略，并可与其他试剂联合用于SSCs的富集。
Spermatogonial stem cells (SSCs) comprise a small population of germ cells with self-renewal potential. Previous studies have shown that SSCs share several common features with stem cells in other self-renewing tissues, including surface markers and proliferative machinery. However, studies of SSCs are severely handicapped by the small number of SSCs and the lack of SSC-specific markers. In the present study, we examined the utility of CDy1 and Rh123, both of which are used for the collection of stem cells in several self-renewing tissues. CDy1 stained germline stem (GS) cells, cultured spermatogonia enriched for SSC activity, after in vitro incubation without exerting toxic effects. Unlike previously reported stem cell-specific dyes, CDy1 was also useful for enrichment of SSCs in both GS cell culture and mature adult testes. Spermatogonial transplantation showed that ∼1 in 66.7 cells exhibited SSC activity after CDH1-based magnetic cell selection and CDy1 staining. In contrast, although Rh123 was previously used successfully to collect SSCs from cryptorchid testes, it was not possible to recover SSCs from both GS cell cultures and wild-type testes. Thus, CDy1 staining will provide a useful strategy for the enrichment of SSCs and may be used in conjunction with other reagents for the enrichment of SSCs.