Reconstitution of Mouse Spermatogonial Stem Cell Niches in Culture
Reconstitution of Mouse Spermatogonial Stem Cell Niches in Culture
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DOI：
10.1016/j.stem.2012.06.011
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发表时间：
2012-10-05
期刊：
影响因子：
23.9
通讯作者：
Shinohara, Takashi
中科院分区：
文献类型：
--
作者：
Kanatsu-Shinohara, Mito;Inoue, Kimiko;Shinohara, Takashi
精原干细胞（SSC）位于曲细精管内的特定龛位。这些小生境被认为是分泌SSC的趋化因子，因为SSC在移植时迁移到它们。然而，这些趋化分子的身份仍然未知。本研究建立了睾丸饲养细胞培养体系，并利用该体系对SSC趋化因子进行了鉴定。当接种在不育小鼠的睾丸细胞上时，SSC迁移到支持细胞下方，形成鹅卵石外观的集落，与造血干细胞产生的集落非常相似。培养的细胞保持SSC活性和生育力至少5个月。鹅卵石集落形成依赖于GDNF和CXCL12，显性阴性GDNF受体转染或CXCL12受体缺陷减少SSC定植。此外，GDNF上调CXCL12受体表达，CXCL12转染Sertoli细胞增加归巢效率。总之，我们的研究结果确定GDNF和CXCL12作为SSC在体外和体内的趋化因子。
Spermatogonial stem cells (SSCs) reside in specific niches within seminiferous tubules. These niches are thought to secrete chemotactic factors for SSCs, because SSCs migrate to them upon transplantation. However, the identity of these chemotactic molecules remains unknown. Here, we established a testis feeder cell culture system and used it to identify SSC chemotactic factors. When seeded on testis cells from infertile mice, SSCs migrated beneath the Sertoli cells and formed colonies with a cobblestone appearance that were very similar to those produced by hematopoietic stem cells. Cultured cells maintained SSC activity and fertility for at least 5 months. Cobblestone colony formation depended on GDNF and CXCL12, and dominant-negative GDNF receptor transfection or CXCL12 receptor deficiency reduced SSC colonization. Moreover, GDNF upregulated CXCL12 receptor expression, and CXCL12 transfection in Sertoli cells increased homing efficiency. Overall, our findings identify GDNF and CXCL12 as SSC chemotactic factors in vitro and in vivo.