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. Conclusion: This study provides a basis for culturing SSCs with gdnf-transfected Sertoli cells as the feeder layer.