中文摘要：

目的探讨大鼠骨髓间充质干细胞能否分化为胆碱能样神经元。方法分离纯化大鼠骨髓间充质干细胞（MSCs）,取2～4代MSCs,进行诱导其向胆碱能神经元分化。诱导方案：10%胎牛血清诱导2d;换液为DMEM/F12培养基,加入碱性成纤维细胞生长因子（bFGF）、巯基乙醇（ME）、维甲酸（RA）、神经生长因子（NGF）,诱导8d;加入表皮生长因子（EGF）,肝素（Heparin）继续诱导8d。空白对照组不作任何处理。观察诱导后的形态变化。采用RT-PCR检测巢蛋白（nestin）,核受体相关因子（Nurr1）,胆碱乙酰转移酶（ChAT）mRNA的表达,间接免疫荧光法检测表达nestin、ChAT、神经核蛋白（NeuN）,乙酰胆碱酯酶（AchE）的阳性细胞。结果诱导后细胞形态逐渐由长梭形变为圆形或椭圆形,突起形成;RT-PCR显示,诱导后的细胞高表达nestin、Nurr1、ChAT,且基因表达相对强度与未诱导的MSCs比较有显著差异（P〈0.01）。免疫荧光法检测显示,诱导后的细胞高表达nestin、ChAT、NeuN、AchE,且阳性细胞大于80%,而对照组除ChAT阳性细胞为1%外,其他均为阴性。结论大鼠MSCs可诱导为胆碱能样神经细胞。

英文摘要：

Objective To explore the differentiation of rat marrow mesenchymal stem cells （MSCs） into cholinergic like-neurons by inducing reagents. Methods MSCs from rat bone marrow were isolated and purified by cell culture,which were induced and differentiated into cholinergic like-neurons with 2~4 generations of MSCs by inducing reagents. The protocol was as follows：MSCs were incubated with 10% FBS for 2 days and then medium was replaced by DMEM/F12 complemented with 10 μg/L of bFGF,0.1 mmol/L of 2-ME,1 mmol/L of RA,10 μg/L of NGF for 8 days. And then 10 μg/L of EGF and 1 250 U/ml of heparin were added into the culture system for culture for 8 days. RT-PCR and gel electrophoresis were used to detect gene expression of nestin,Nurr-1,choline acetyltransferase （CHAT）. Immunological fluorescent staining was used to measure nestin,CHAT,NeuN,acetylcholine esterase （AchE） positive cells. Results Induced cells gradually became from long shuttle into round or ellipse with neuron-like dendrite. RT-PCR showed that the induced neuron-like cells expressed special neurocyte genes of nestin,Nurron,CHAT. Indirect immunological fluorescent staining showed positive cells with nestin,CHAT,NeuN,AchE reached 80% respectively. Conclusions MSCs from rat bone marrow can differentiate into cholinergic like-neurons.