中文摘要：

目的：从小鼠睾丸中克隆胶质细胞源神经营养因子基因gdnf,构建真核表达载体,并转染支持细胞,以便用作培养精原干细胞（SSCs）的滋养层。方法：以正常成年昆明鼠为材料,提取小鼠睾丸组织中总RNA后,以RT-PCR技术克隆小鼠睾丸gdnf基因,构建真核表达载体,并转染TM4细胞（睾丸支持细胞株）,在转染后40h进行免疫荧光鉴定。结果：成功克隆小鼠睾丸gdnf基因的cDNA,测序正确,免疫荧光细胞染色显示转染后的支持细胞中有GDNF蛋白表达。结论：本研究为以转染了gdnf基因的支持细胞作饲养层培养SSCs奠定了基础。

英文摘要：

Objective： To clone the glial cell line-derived neurotrophic factor （GDNF） from the mouse testis, construct the eukaryotic expression vector and transfect this vector into Sertoli cells in order to use the gdnf-transfected Sertoli cells as the feeder layer to cultivate spermatogonial stem cells （SSCs）. Methods ： Total RNA was extracted from the testes of normal mature mice and gdnf was cloned and amplified using RT-PCR, inserted into the eukaryotic expression vector and transfected into sertoli cells （TM4 cell line）. Immunofluorescence with anti-GDNF antibodies was performed at 40 h following the transfection. Results ： gdnf cDNA was cloned successfully, and GDNF expressed after transfected into Sertoli cells. Conclusion ： This study provides a basis for culturing SSCs with gdnf-transfected Sertoli cells as the feeder layer.