人骨髓间质干细胞定向分化为神经元样细胞的实验  

Experiment on orientating differentiation of human mesenchymal stem cells into neuron-like cells

作　　者：杨晓苏[1] 吴海香[1] 肖波[1] 邓益东[1] 

机构地区：[1]中南大学湘雅医院神经内科,湖南省长沙市410008

出　　处：《中国临床康复》2005年第46期9-11,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(30170330);湖南省自然科学基金资助项目(02JJY3016)~~

摘　　要：目的:体外定向诱导人骨髓间质干细胞分化为神经元样细胞并观察其传导功能。方法:实验于2003-06/2004-02在中南大学湘雅医院完成。采用淋巴细胞分离液(1.007g/L)梯度离心分离人骨髓间质干细胞,体外进行培养扩增,丁化羟基苯甲醚和二甲基亚砜诱导人骨髓间质干细胞分化为神经元样细胞,免疫细胞化学方法鉴定神经元特异性标志物,神经元特异性烯醇化酶、神经丝蛋白,突触素蛋白,胶质细胞标志物胶质纤维酸性蛋白。结果:加入诱导剂3h后,大多数细胞转变为类似于神经元的形态,有长的突起,相互之间连接呈网状,并表达神经元特异性烯醇化酶,阳性率为(75.0±6.5)%、神经丝蛋白阳性率为(68.0±4.2)%及胶质纤维酸性蛋白阳性率为(25.0±6.4)%,但胶质纤维酸性蛋白表达明显较前两者弱(P<0.05)。神经元细胞胞体表达突触素蛋白,突起未见表达。结论:人骨髓间质干细胞在体外可以分化为神经元样细胞;神经元样细胞其突起不具备传导功能。AIM： To investigate the differentiation of human mesenchymal stem cells （hMSCs） into neuron-like cells in vitro and explore the function of their conduction. METHODS： The experiment was conducted at the Xiangya Hospital, Central South University between June 2003 and February 2004. The hMSCs were separated with gradient centrifugalization with lymphocyte separating medium （1.007 g/L）, and expanded in culture medium in vitro. The hMSCs were induced to differentiate into neuron-like ceils with butylated hydroxyanisole （BHA） and dimethylsulfoxide （DMSO）. Neuron specific enolase （NSE）, neurofilament protein, synaptophysin protein and glial fibrillary acidic protein （GFAP） were detected by immunohistochemistry in neuron-like cells. RESULTS： Most of the hMSCs differentiate into neuron-like cells after being introduced for 3 hours. Neuron-like cells had long processes contacting with each other. Neuron-like cells expressed the neuronal marker NSE with the （75.0±6.5）% positive rate, neurofilament protein with the （68.0±4.2）% positive rate and GFAP with the （25.0±6.4）% positive rate, but the expression of GFAP was weaker than that of the former two （P 〈 0.05）. The neuronal body expressed synaptophysin protein, and there was no expression in processus. CONCLUSION： The hMSCs are able to differentiate into neuron-like cells in vitro, but its processus do not have the function of conduction.

关 键 词：骨髓细胞 干细胞 细胞分化 神经元 

分 类 号：R394.2[医药卫生—医学遗传学]