Conservation of spermatogonial stem cell self-renewal signaling between mouse and rat
Conservation of spermatogonial stem cell self-renewal signaling between mouse and rat
复制标题
DOI：
10.1073/pnas.0506970102
复制
发表时间：
2005-10-04
影响因子：
11.1
通讯作者：
Brinster, RL
中科院分区：
文献类型：
--
作者：
Ryu, BY;Kubota, H;Brinster, RL
精原干细胞（SSCs）的自我更新是维持精子发生和物种延续的基础。干细胞自我更新的分子机制是干细胞生物学中的一个基本问题。最近，我们确定了小鼠精原干细胞自我更新所必需的生长因子，并建立了一个无血清培养体系，使其在体外增殖。为了确定SSC复制的刺激信号是否在不同物种之间是保守的，我们将培养系统扩展到大鼠SSC。最初，通过使用流式细胞术分析开发了一种评估SSC体外扩增的方法，随后，我们发现胶质细胞系衍生的神经营养因子、可溶性胶质细胞系衍生的神经营养因子家族受体α-1和碱性成纤维细胞生长因子的组合支持大鼠SSC的增殖。当与三种因子一起培养时，干细胞连续增殖> 7个月，并且将培养的SSC移植到受体大鼠中产生供体干细胞衍生的后代，表明培养的干细胞是正常的。大鼠SSC复制所需的生长因子与小鼠相同;因此，SSC自我更新的信号传导因子在这两个物种中是保守的。由于许多哺乳动物（包括人类）的SSC在移植后可以在小鼠曲细精管中复制，因此SSC自我更新所需的生长因子在许多不同物种中可能是保守的。此外，大鼠精原干细胞长期培养体系的建立为基因打靶技术进行大鼠生殖系修饰奠定了基础。
Self -renewal of spermatogonial stem cells (SSCs) is the foundation for maintenance of spermatogenesis throughout life in males and for continuation of a species. The molecular mechanism underlying stem cell self-renewal is a fundamental question in stem cell biology. Recently, we identified growth factors necessary for self-renewal of mouse SSCs and established a serum-free culture system for their proliferation in vitro. To determine whether the stimulatory signals for SSC replication are conserved among different species, we extended the culture system to rat SSCs. Initially, a method to assess in vitro expansion of SSCs was developed by using flow cytometric analysis, and, subsequently, we found that a combination of glial cell line-derived neurotrophic factor, soluble glial cell line-derived neurotrophic factor-family receptor alpha-1 and basic fibroblast growth factor supports proliferation of rat SSCs. When cultured with the three factors, stem cells proliferated continuously for > 7 months, and transplantation of the cultured SSCs to recipient rats generated donor stem cell-derived progeny, demonstrating that the cultured stem cells are normal. The growth factor requirement for replication of rat SSCs is identical to that of mouse; therefore, the signaling factors for SSC self-renewal are conserved in these two species. Because SSCs from many mammals, including human, can replicate in mouse seminiferous tubules after transplantation, the growth factors required for SSC self-renewal may be conserved among many different species. Furthermore, development of a long-term culture system for rat SSCs has established a foundation for germ-line modification of the rat by gene targeting technology.