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目的:观察嗅鞘细胞(OECs)诱导神经干细胞(NSCs)分化的方向,分化后的细胞存活和电活动情况。方法:将分离培养人胚OECs细胞经纯化后接种于培养皿内,2d后加入神经干细胞,设置5×10-6RTRA组,5%FBS对照组,分别于第10天后行NSE免疫组化检测,于7d、14d、21d对嗅鞘细胞组和RA组行tunnel细胞凋亡检测,于12d行膜片钳检测。结果:NSE阳性率OECs组80.3%,RTRA组77.5%,FBS组2%。7d、14d、21d对嗅鞘细胞组和RTRA组tunnel细胞凋亡比例分别为0.9%、1.2%、1.4%,1%、3.3%、8.6%,分化后的神经元记录到快速激活快速失活内向钠电流及慢激活慢失活能延迟整流性外向钾电流。结论:OECs能诱导NSCs向神经元方向分化,促进分化后的细胞存活,并且有神经元样电活动,因此NSCs分化后的细胞具备了替代受损的神经元细胞的功能。
OBJECTIVE: To observe the direction of differentiation of neural stem cells (NSCs) induced by OECs and the cell survival and electrical activity after differentiation. Methods: The cultured human embryonic OECs cells were purified and seeded into petri dishes. After 2 days, neural stem cells were added and 5 × 10-6 RTRA group and 5% FBS control group were set up, and NSE immunohistochemistry was performed on the 10th day respectively. At 7d, 14d, 21d, olfactory ensheathing cells group and RA group were tested for apoptosis of tunnel cells, and patch clamp was performed on the 12th day. Results: The positive rate of NSE was 80.3% in OECs group, 77.5% in RTRA group and 2% in FBS group. The percentage of apoptotic cells in the olfactory ensheathing cell group and the RTRA group was 0.9%, 1.2%, 1.4%, 1%, 3.3% and 8.6% at 7d, 14d and 21d respectively. The differentiated neurons were rapidly inactivated Inward sodium current and slow activation slow inactivation delay rectifier outward potassium current. CONCLUSIONS: OECs can induce the differentiation of NSCs to neurons, promote the survival of differentiated cells and have neuronal-like electrical activity. Therefore, the differentiated NSCs have the function of replacing damaged neurons.