A recombinant retrovirus vector containing the glial cell line-derived neurotrophic factor (GDNF) gene was constructed and transfected into Schwann cells (SCs) to investigate the possibility of GDNF transfection and functional expression of transfected SCs, including GDNF secretion and its mRNA expression. We found that transfection of the GDNF gene into SCs led to significantly enhanced expression of GDNF mRNA. The rate of GDNF secretion by GDNF-SCs was also increased. Functionally, more surviving motoneurons were seen when they were cocultured in GDNF-SC-conditioned medium than when they were in normal SC-conditioned medium. When bridging a rat sciatic nerve defect with a conduit filled with GDNF-transfected SCs, nerve regeneration was better than that of the control. In conclusion, transfection of SCs with the GDNF gene could enhance SC function. Application of genetically modified SCs could be a better way to promote nerve regeneration.