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[目的]通过体外共培养观察人嗅鞘细胞(OECs)对骨髓间充质干细胞(MSCs)增殖分化的影响。[方法]取5个月以上引产胎儿嗅球及骨髓,分离培养及鉴定OECs及MSCs,携带绿色荧光蛋白(GFP)基因的腺相关病毒转染标记0ECs,双苯亚甲胺荧光染料标记MSCs,将两种细胞按1:1比例进行共培养,MSCs单独培养组为阴性对照组,免疫细胞化学方法检测MSCs表达巢蛋白(nestin)、神经元特异性核蛋白(NeuN)、神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF-M)、胶质纤维酸性蛋白(GFAP)情况。[结果]MOI值为1×106时0ECs表达GFP阳性率为84.89%,对细胞生长活性无明显影响。共培养时两种细胞生长良好,共培养7 d未见形态学改变,经复方丹参注射液诱导后部分MSCs呈现神经元样改变,表达nestin、NSE、NeuN、NF-M、GFAP特异性标志,且共培养组的分化率明显高于单独MSCs培养组,有显著性差异。[结论]OECs可通过分泌可溶性物质及细胞之间的相互作用启动MSCs分化程序,提高MSCs向神经细胞分化率。
[Objective] To observe the effects of OECs on the proliferation and differentiation of bone marrow mesenchymal stem cells (MSCs) by co-culture in vitro. [Methods] Fetal olfactory bulb and bone marrow were induced in fetus for more than 5 months. OECs and MSCs were isolated and cultured. Adeno-associated virus carrying green fluorescent protein (GFP) gene was transfected into 0ECs and labeled with MSCs. The two kinds of cells were co-cultured in a ratio of 1: 1. MSCs cultured alone were negative control group. Immunocytochemistry was used to detect the expression of nestin, neuron specific nucleoprotein (NeuN), neuron specific enolase Enzyme (NSE), neurofilament (NF-M), glial fibrillary acidic protein (GFAP) situation. [Results] The positive rate of GFP expression in OECs was 84.89% at the MOI value of 1 × 106, and had no significant effect on the cell growth activity. After co-cultured for 7 days, no morphological changes were observed. After induced by compound Danshen injection, some MSCs showed neuron-like changes, and expressed nestin, NSE, NeuN, NF-M and GFAP specific markers, And the differentiation rate of co-culture group was significantly higher than that of MSCs alone group, there was a significant difference. [Conclusion] OECs can initiate MSCs differentiation process through the secretion of soluble substances and the interaction between cells, and improve the differentiation rate of MSCs to nerve cells.