Spermatogonial stem cell enrichment by multiparameter selection of mouse testis cells
Spermatogonial stem cell enrichment by multiparameter selection of mouse testis cells
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DOI：
10.1073/pnas.97.15.8346
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发表时间：
2000-07-18
影响因子：
11.1
通讯作者：
Brinster, RL
中科院分区：
文献类型：
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作者：
Shinohara, T;Orwig, KE;Brinster, RL
精原干细胞在睾丸中启动并维持精子的发生。为了发挥这一作用，干细胞必须自我复制并产生子细胞，子细胞可以扩大和分化形成精子。尽管精原干细胞在男性生殖中起着重要的作用，但人们对其形态和生化特征知之甚少。这部分是由于精原干细胞在睾丸中是一种极其罕见的细胞群，而富集它们的技术才刚刚开始建立。在这项研究中，我们采用了多参数选择策略，将体内隐睾模型与体外荧光激活细胞分选分析相结合。基于光散射特性和细胞表面分子α 6-整联素、α v-整联素的表达，采用荧光激活细胞分选方法对睾丸隐睾细胞进行分离。c-kit受体。从这些分析中得出了两个重要的观察结果。第一。来自成年隐睾的精原干细胞很少或不表达c-kit。第二。在这项研究中，最有效的富集策略。选择具有低侧散射光散射特性的细胞。α 6-整合素阳性染色，α v-整合素阴性或低表达，导致精原干细胞富集166倍。这些特征的识别将有助于进一步纯化这些有价值的细胞，并促进对控制精原干细胞自我更新和分层分化的分子机制的研究。
The spermatogonial stem cell initiates and maintains spermatogenesis in the testis. To perform this role, the stem cell must self replicate as well as produce daughter cells that can expand and differentiate to form spermatozoa. Despite the central importance of the spermatogonial stem cell to male reproduction, little is known about its morphological or biochemical characteristics. This results, in part, from the fact that spermatogonial stem cells are an extremely rare cell population in the testis, and techniques for their enrichment are just beginning to be established. In this investigation, we used a multiparameter selection strategy, combining the in vivo cryptorchid testis model with in vitro fluorescence-activated cell sorting analysis. Cryptorchid testis cells were fractionated by fluorescence-activated cell sorting analysis based on light-scattering properties and expression of the cell surface molecules alpha 6-integrin, alpha v-integrin. and the c-kit receptor. Two important observations emerged from these analyses. First. spermatogonial stem cells from the adult cryptorchid testis express little or no c-kit. Second. the most effective enrichment strategy, in this study. selected cells with low side scatter light-scattering properties. positive staining for alpha 6-integrin, and negative or low alpha v-integrin expression, and resulted in a 166-fold enrichment of spermatogonial stem cells. Identification of these characteristics will allow further purification of these valuable cells and facilitate the investigation of molecular mechanisms governing spermatogonial stem cell self renewal and hierarchical differentiation.