摘要
目的 探索成人骨髓间充质干细胞在体外定向分化为神经元样细胞的条件。方法 采用Ficoll paque( 1 0 77g/ml)分离液密度梯度离心从人的骨髓分离出骨髓间充质干细胞 ,体外扩增到第 10代 ,选择 4组不同组合的神经分化诱导条件 ,分别诱导骨髓间充质干细胞向神经元细胞分化 ,通过免疫细胞化学鉴定诱导后细胞神经元烯醇化酶、神经丝蛋白、胶质纤维酸性蛋白的表达情况。结果 各组诱导剂诱导后 ,骨髓间充质干细胞分化为具有典型的神经元形态的细胞 ,免疫细胞化学显示、NF M呈强阳性反应 ,NSE表达强度提高 ,在最佳的分化条件下 ,即加入bFGF、ATRA、BME、BHA、DMSO诱导条件下 ,大约 80 %细胞表达NF M ,86%的细胞为NSE强阳性。结论 成人MSCs在体外可定向诱导分化为神经元样细胞 。
Objective To investigate the conditions of the oriented differentiation of human adult mesenchymal stem cells(MSCs) into neuron like cells in vitro . Methods MSCs were isolated from human adult bone marrow through density gradient with Ficollll paque(1.077 g/ml) and then amplified as undifferentiated cells for 10 generations. Four different groups of neural differentiation inducing conditions were selected for the induction of MSCs differentiation into neuronal cells, respectively. The expressions of neuron specific enolase(NSE), neurofilament protein(NF M) and glial fibriliary acidic protein(GFAP) were detected by immunocytochemistry. Results MSCs were induced to differentiate into cells with a typical neuronal morphology, expressing NF M and increased levels of NSE. The induced neuron like cells extended long processes and filopodia. Under an optimal differentiation condition, i.e. after treatment with ATRA, bFGF, BME and DMSO, almost 80% of the cells expressed NF M and 86% of the cells expressed intensively NSE. Conclusion MSCs can be induced to differentiate into neuron like cells with high positive rate of induced differentiation.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2003年第11期941-944,共4页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 30 0 70 382 )
高等学校骨干教师资助项目 (教技司 2 0 0 0 - 0 6 )
重庆市教委科学基金资助项目 ( 99-0 0 0 1)
关键词
骨髓间充质干细胞
神经分化
mesenchymal stem cells
neural differentiation
