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BACKGROUND: Previous studies have demonstrated that the combination of olfactory ensheathing cells (OECs) and neurotrophic factor-3 (NT-3) in the rat lateral ventricle can promote nerve axonal regeneration and myelin sheath repair. However, this effect remains very short-lived.OBJECTIVE: To transfect NT-3 into OECs and to observe the biological activity of OEC-expressing NT-3.DESIGN, TIME AND SETI'ING: This genetic engineering, in vitro experiment was performed in the Provincial Hospital Affiliated to Shandong University between January 2007 and October 2008.MATERIALS: Trizol Reagent kit was purchased from Gibco, USA; reverse transcription kit, NT-3Emax lmmunoAssay System reagent was purchased from Promega, USA.METHODS: Neonatal Wistar rat OECs were established as primary cultures and were transfected with pN2A-NT-3 viral vector. The OECs with the highest virus titer and stable cellular growth served as the transfection group; OECs transfected with NT-3-free retrovirus carrier pN2A served as theempty vector group; un-transfected OECs served as the control group. After adherence, the logarithmically cultured PC12-TrkC cells were plated in OECs supernatant from the transfectJon and empty vector groups, as well as 20 μL PBS, and cultured for 4 days.MAIN OUTCOME MEASURES: NT-3 mRNA expression in OECs, fluorescence of NT-3-positivecells in the transfection group and control group; influence of OECs secreting NT-3 on the differentiation ratio of PC12-TrkC cells.RESULTS: NT-3 mRNA expression was observed 24 hours after transfeotion and lasted for 28 days,which was greater than the control and empty vector groups (P