Objective To explore the biocompatibility of rat olfactory ensheathing cells(OECs) with electrospun nanofiber-PLGA (poly[D,L-lactide-co-glycolide]) membrane. Methods The OECs of adult rats were seeded on the electrospun nanofiber-PLGA membrane. The morphology and adhesion to the membrane of OECs were observed by phase contrast microscopy.The proliferation ability of OECs was evaluated by CFDA SE immunofluorescent at 1-5 days after seeding. The control group was seeded into poly-L-lysine-treated plate. Moreover, the electrospun nanofiber-PLGA membrane was also implanted in rats to observe the biocompatibility. Results The purity of OECs obtained from in vitro culture was greater than 90%.The microscopic findings showed that the OECs grew well on the electrospun nanofiber-PLGA membrane, with good adhesion ability and good cell state; the cell also had an orientation growth along the nanofibers.The OECs had a normal proliferation in the electrospun nanofiber-PLGA membrane at 1-5 days after seeding, and the cell number was not significantly different from that of the control group. There was no death in rats after the PLGA film was transplanted, and the PLGA membrane was fused with the surrounding tissues, with some degradation at the fusion sites with the tissues. Conclusion The electrospun nanofiber-PLGA membrane shows a satisfactory biocompatibility, and OECs have good adhesion and proliferation on it, indicating that the membrane might be a potential tissue engineering nerve graft when combined with OECs.