睾丸支持细胞的基本生物学特性  

Basic biological characteristics of Sertoli cells

作　　者：韩妲丽[1] 王晓云[1] 邢新[1] 周广东[2] 刘伟[2] 曹谊林[2] 

机构地区：[1]第二军医大学附属上海市长海医院整形外科,上海200433 [2]上海交通大学医学院附属第九人民医院组织工程实验室,上海200011

出　　处：《生殖医学杂志》2008年第5期361-365,共5页Journal of Reproductive Medicine

基　　金：国家973计划课题项目(2005CB522705-4);国家自然科学基金资助项目(30600619);国家博士后科研基金资助项目(2005037461)

摘　　要：目的通过了解睾丸支持细胞(Sertoli cells,SC)的基本生物学特性,探讨SC鉴定的良好方法。方法联合应用复合胶原酶及差速贴壁法从睾丸组织中分离、培养SC,光镜和电镜下观察细胞的形态、MTT法测定细胞的生长曲线,观察其在体外培养条件下的增殖特性;利用免疫细胞染色及免疫荧光染色的方法,检测Fas配体(FasL)的表达,观察其免疫功能;应用吖啶橙荧光染色进行细胞鉴定。结果联合应用复合胶原酶及差速贴壁法分离、培养的细胞在光镜下呈长柱状及三角形,增殖能力强,电镜下胞核中可见特异性卫星小体,胞质中细胞器丰富,免疫细胞染色及免疫荧光染色证实其高表达FasL,吖啶橙荧光染色可见胞核中含有大量异染色质,核仁明显,证实其为SC。结论复合胶原酶及差速贴壁法分离的SC具有良好的增殖能力及免疫功能,电镜、吖啶橙荧光染色是鉴定SC方便、有效的方法。Objectives： Sertoli cells can induce immune privilege. When co-transplanted with Sertoli cells, allograft or xenotransplanted cells can survive longer. But the identification of sertoli cells after co- transplantation was not full established. The basic biological characteristics of Sertoli cells was studied here, hopefully contributing to the identification of them. Methods： Sertoli cells from human testis were isolated by the technique of eollagenase digestion and different-speed adherence. The morphology of Sertoli cells was observed under light microscope and electron microscope. The growth curve was measured by MTT method. The expression of FasL in Sertoli cells was detected by immunocytochemistry and fluorescence staining. Sertoli cells were identified by acridine orange stain. Results： Abundant Sertoli cells can be obtained from the human testis by collagenase digestion and different-speed adherence method. They had the appearance of long pillar or triangle and showed a high proliferation rate. Specific satellite globules in nucleus and abundant organelles in cytoplasm were observed under electron microscope. Sertoli cells expressed FasL and had abundant heterochromatin and limpid nucleolus with acridine orange fluorescence stain.Conclusions： Abundant Sertoli cells with proliferative activity and immune function can be harvested by collagenase digestion and different-speed adherence methods from the human testis. It is potential to identify Sertoli cells by electron microscope and acridine orange staining.

关 键 词：支持细胞 FAS配体 吖啶橙 

分 类 号：R588[医药卫生—内分泌] R617[医药卫生—内科学]