Cloning of gdnf in the mouse testis and its expression in sertoli cells.
- Author:
Cui-mi DUAN
1
;
En-zhong LI
;
Shi-qing ZHANG
;
Chang-yong WANG
;
De-xue LI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cloning, Molecular; Gene Expression; Glial Cell Line-Derived Neurotrophic Factor; genetics; Male; Mice; Mice, Inbred Strains; RNA; genetics; metabolism; Reverse Transcriptase Polymerase Chain Reaction; Sertoli Cells; metabolism; Testis; cytology; metabolism; Transfection
- From: National Journal of Andrology 2007;13(11):975-978
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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).
METHODSTotal 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.
RESULTSgdnf cDNA was cloned successfully, and GDNF expressed after transfected into Sertoli cells.
CONCLUSIONThis study provides a basis for culturing SSCs with gdnf-transfected Sertoli cells as the feeder layer.
