摘要
[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 effect of OECs on the generation and differentiation of MSCs and investigate the potential mechanism. [ Method ] Olfactory bulbs and human bone marrow were got from 5 months old induced labor fetus to sepa- rate and culture OECs and MSCs. OECs were transfected by recombinant adeno-associated virus carrying the GFP gene and MSCs＇ nuclei were labeled with Hoechst dye. Then labeled OECs and MSCs were co-cultured in vitro with the proportion of one to one. MSCs cultured alone were treated as a control group. Neuron-specific enolase （ NSE）, neurofila-ment （ NF）, nestin, glial fibrillary acidic protein （GFAP） were detected by immunocytochemistry to observe the expressing of MSCs. [ Result] 84. 89% of OECs expressed GFP,which had no effect on the activity of cells ,when MOI was 1 ~ 106. Both kinds of cells grew well and no significant contact inhabitation was observed in 7 days. After induced by the DMEM/Danshen injection, part of MSCs could transformed into neuron-like cells and expressed some special proteins such as nestin, NSE, NeuN and NF-M. The trans- formation percentage of the combined transplantation group was higher than that of single MSCs group and the difference was sig- nificant. [ Conclusion] OECs can active MSCs＇ differentiation program by some soluble material and contact between cells, so that the transformation from MSCs to neuron like cells is more efficiently.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2013年第15期1562-1568,共7页
Orthopedic Journal of China
基金
广东省科技计划项目(编号:2004A30201002)
关键词
嗅鞘细胞
骨髓间充质干细胞
绿色荧光蛋白
共培养
olfactory ensheathing cells, bone marrow mesenchymal stem cells, green fluorescent protein, co-culture
