组织工程与重建外科杂志2011，Vol.7Issue(4)：186-190,206,6.DOI:10.3969/j.issn.1673-0364.2011.04.002
睾丸Leydig干细胞分离、纯化及鉴定的体外研究
Research of Leydig Stem Cells: Separation, Purification and Identification In Vitro
摘要
Abstract
Objective To improve the methods for separation and purification of Leydig stem cells (SLCs) and observe its biological characteristics and to identify the cells. Methods Leydig stem cells were gained from the testes of Wistar rats by ways of collagenase digestion, differential adhesion and two-antibodies magnetic activated cell sorting. And then the cells were cultured in modified culture media and its reproductive activity was tested by cell counting kit (CCK) method. SLCs were identified by immunohistochemistry (IHC) staining with the detection of PDGFRa, LIFR and 3β-HSD. Testosterone level was detected by RIA method after SLCs were induced for differentiation. Results About 83 000 cells were available in each testis and the PDGFRα+ SLCs could reach to 98% by IHC staining. SLCs proliferated actively in modified culture media and could maintain undifferentiated state within a month in vitro. IHC staining results showed that the expression of PDGFRa and LIFR were positive and the expression of 3|$-HSD was negative. Testosterone secretion could be detected at day 5 while SLCs were cultured in media added with T3 and HCG. 3β-HSD of differentiated cells turned to be positive by IHC staining. Conclusion PDGFRa VLHR' cells could be obtained by differential adhesion and magnetic activated cell sorting method. The cells were in accordance with the characteristics necessary for SLCs. This method has the advantages of simple, small cell damage and high purity.关键词
Leydig干细胞/磁性活化细胞分选/细胞培养/分化/鉴定Key words
Leydig stem cells (SLCs)/MACS/Cell culture/Differentiation/Identification分类
生物科学引用本文复制引用
肖斌,王晓云,周广东,周英晋,毕宏达,朱吉,张敬德,邢新..睾丸Leydig干细胞分离、纯化及鉴定的体外研究[J].组织工程与重建外科杂志,2011,7(4):186-190,206,6.基金项目
国家自然科学基金(30970736). （30970736）
