温度及BRL饲细胞对体外小鼠精原干细胞的影响  

The Effects of Temperature and BRL Feeders on Mouse Spermatogonial Stem Cells in Vitro

作　　者：李莲军[1] 苗永旺[1] 程美玲[1] 李卫真[1] 卢克焕[2] 

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201 [2]广西大学动物繁殖研究所,广西南宁530005

出　　处：《云南农业大学学报》2004年第5期580-583,共4页Journal of Yunnan Agricultural University

基　　金：云南省自然科学基金(2003C0046M);广西自然科学基金(桂科自0339005)。

摘　　要：将6日龄小鼠生精上皮单细胞分别接种于BRL和STO饲养层上,于32℃或37℃培养,研究其对精原干细胞的影响。结果:在培养的第1周内,2个及4个相连的精原细胞合胞体明显增多。培养1周后,多数精原细胞经短暂的存活及分裂活动后退化消失,培养体系中仅保留下少量单个及由细胞间桥相连的双个及4个成串或成团的A型精原细胞。这些细胞在随后的培养过程中,不表现明显的分裂活动,呈碱性磷酸酶阳性反应。根据精原干细胞生物学特性,它们极有可能就是精原干细胞及其子代细胞。不同条件培养体系中的精原干细胞的生物学行为无明显不同,培养60d时均仅有少量精原干细胞存活。结论:BRL细胞能用作饲养层促进精原干细胞存活,但对其更新性增殖没有明显作用;32～37℃对精原干细胞的生物学行为无明显影响,均可用于培养精原干细胞。The seminiferous epithelial cells dissociated from 6 days postpartum mice were cultured on buffalo rat liver (BRL) cell feeders or on STO feeders at 32℃ or at 37℃respectively to study the effects of BRL feeders and different temperatures on culture of mouse spermatogonial stem cells. During the first week of culture, the spermatogonial syncytia of 2-and 4-cell increased obviously. After a week of culture only some single, paired, aligned or clustered spermatogonia maintained. They showed no obvious proliferation or differentiation in subsequent culture. These cells were spherical and expressed AP activity. They were most likely spermatogonial stem cells and their first several progenies according to the biological characteristics of spermatogonial stem cells. Under different culture conditions mentioned above, the biological behaviors of spermatogonial stem cells showed no obvious difference and only small number of spermatogonial stem cells remained when they were cultured for 60 days. It is concluded that BRL cells can be used as feeders to promote survival but not renewal of spermatogonial stem cells. The temperatures from 32℃ to 37℃ have on special effects on spermatogonial stem cells and can be used for culture of them.

关 键 词：精原干细胞 BRL饲养层 温度 体外培养 小鼠 

分 类 号：Q954.43[生物学—动物学]