[小鼠睾丸中胶质细胞源性神经营养因子的克隆及其在支持细胞中的表达]
[Cloning of gdnf in the mouse testis and its expression in sertoli cells].
作者信息
Duan Cui-mi, Li En-zhong, Zhang Shi-qing, Wang Chang-yong, Li De-xue
机构信息
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
出版信息
Zhonghua Nan Ke Xue. 2007 Nov;13(11):975-8.
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.
目的
从小鼠睾丸中克隆胶质细胞源性神经营养因子（GDNF），构建真核表达载体并将其转染至支持细胞，以便将转染了gdnf的支持细胞用作饲养层来培养精原干细胞（SSCs）。
方法
从正常成熟小鼠的睾丸中提取总RNA，采用RT-PCR克隆并扩增gdnf，将其插入真核表达载体并转染至支持细胞（TM4细胞系）。转染后40小时用抗GDNF抗体进行免疫荧光检测。
结果
成功克隆出gdnf cDNA，转染至支持细胞后GDNF得以表达。
结论
本研究为以转染了gdnf的支持细胞作为饲养层培养精原干细胞提供了依据。
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