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目的探索体外嗅鞘细胞对神经干细胞增殖的影响及其机制。方法体外培养神经干细胞和嗅鞘细胞,分别采用共培养及共培养液培养,观察嗅鞘细胞对神经干细胞增殖的影响。用免疫组织化学和RT-PCR检测嗅鞘细胞表达的细胞因子及其受体的情况。结果共培养及共培养液培养4d后,神经干细胞细胞数量明显增多并有部分细胞分化,与对照组相比有显著性差异(P<0.05)。RT-PCR结果显示,嗅鞘细胞表达-βNGF、BDNF、NT-3、PDGF-Bt、rkA和trkB的mRNA,而不表达NT-4。免疫组织化学嗅鞘细胞呈NGF、BDNF和p75染色阳性。结论嗅鞘细胞表达或分泌大量细胞因子,如NGF、BDNF、NT-3和PDGF,并可能通过旁分泌或靶源性模式作用于神经干细胞,促进其增殖。
Objective To explore the effect of olfactory ensheathing cells on proliferation of neural stem cells in vitro and its mechanism. METHODS: Neural stem cells and olfactory ensheathing cells were cultured in vitro. The co-culture and co-culture medium were used respectively to observe the effects of olfactory ensheathing cells on the proliferation of neural stem cells. The expression of cytokines and their receptors in olfactory ensheathing cells was detected by immunohistochemistry and RT-PCR. Results After co-culture and co-culture for 4 days, the number of neural stem cells was significantly increased and some cells were differentiated, which was significantly different from the control group (P <0.05). RT-PCR results showed that OECs expressed mRNA of -βNGF, BDNF, NT-3, PDGF-Bt, rkA and trkB, but not NT-4. OECs were positive for NGF, BDNF and p75 immunohistochemistry. Conclusion Olfactory ensheathing cells express or secrete a large number of cytokines such as NGF, BDNF, NT-3 and PDGF, and may act on neural stem cells through paracrine or target-derived mode to promote their proliferation.