摘要
精原干细胞具有自我增殖和分化为精子的能力。精原干细胞的培养、移植和体外诱导分化等研究将使我们最终阐明精原干细胞的自我更新机制和精子发生机理。经过一年多的研究,我们不仅建立了用含血清培养基对分离的乳鼠睾丸细胞进行差异贴壁分选来富集生殖细胞的简单方法,而且成功地对分选出的精原干细胞进行了近一个月的无血清培养。流式分析结果表明,差异贴壁分选能将精原干细胞富集11倍以上。移植分析表明,分选出的精原干细胞具有在受体鼠的曲精小管中产生克隆的能力和正常生精作用。免疫细胞化学和RT-PCR检测表明,培养的细胞表达精原干细胞标志基因。该研究为体外长期培养小鼠或其它哺乳动物的精原干细胞提供了一个范例,也为利用基因修饰的精原干细胞通过移植产生转基因动物奠定了基础。
Spermatogonial stem ceils （SSCs） have abilities to self-renew and differentiate into spermatozoa. Studies of culture, transplantation and in vitro induced differentiation of SSCs will help us to ultimately elucidate mechanisms controlling SSC self-renewal and spermatogenesis. Having studied over one year, we not only established a simple method, namely differential adherence selection, to enrich germ ceils from mouse pup testis ceils with serum-containing medium, but had successfully cultured SSCs for about one month with serum-free defined medium. Flow cytometric analysis confirmed that the SSC population could be increased by more than 11 folds. Transplantation assay indicated that the selected SSCs could colonize into recipient seminiferous tubules and restore normal spermatogenesis. Immunocytochemistry and RT-PCR analysis showed that our cultured ceils expressed SSC marker genes. Therefore, our investigation provides a paradigm for long-term culture of SSCs from mouse or other mammals, and establishes the foundation to produce transgenic animals by using gene-modified SSCs.
出处
《潍坊学院学报》
2007年第2期61-66,共6页
Journal of Weifang University
基金
国家自然科学基金(批准号:30300182)资助项目
关键词
小鼠精原干细胞
分离
移植
培养
mouse spermatogonial stem cells, isolation, transplantation, culture
