[Cloning of gdnf in the mouse testis and its expression in sertoli cells].
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.


CONCLUSIONS
This study provides a basis for culturing SSCs with gdnf-transfected Sertoli cells as the feeder layer.
Duan
Cui-mi
CM
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China. duancm2006@yahoo.com.cn
Li
En-zhong
EZ
Zhang
Shi-qing
SQ
Wang
Chang-yong
CY
Li
De-xue
DX
D000818:Animals; D003001:Cloning, Molecular; D015870:Gene Expression; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D012313:RNA / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000378:metabolism*; D013737:Testis / Q000166:cytology / Q000378:metabolism*; D014162:Transfection
D051100:Glial Cell Line-Derived Neurotrophic Factor; D012313:RNA