摘要
目的探讨音速波状蛋白(Shh)促进人骨髓间充质干细胞(MSCs)体外定向分化为多巴胺能神经元样细胞的作用。方法体外分离、扩增和鉴定人骨髓MSCs。采用不同诱导方案诱导MSCs向神经元和多巴胺能神经元样细胞定向转化后,进行抗神经巢蛋白(Nestin)、神经元特异烯醇化酶(NSE)、神经胶质纤维酸性蛋白(GFAP)、酪氨酸羟化酶(TH)和多巴胺转运体(DAT)等免疫细胞化学染色,并计算阳性细胞百分率。结果实验组诱导后MSCs能分化为具有典型神经元形态的细胞,可见NSE、Nestin、GFAP、TH和DAT等神经细胞标志表达;对照组MSCs细胞形态无明显变化,上述特异性标志物表达均为阴性。实验2组(诱导方案含Shh)与1组(诱导方案不含Shh)的NSE、Nestin、GFAP阳性细胞百分率的差异无统计学意义,但实验2组TH和DAT阳性细胞百分率明显高于实验1组,差异具有统计学意义(P<0.05)。结论Shh可促进MSCs分化为多巴胺能神经元样细胞。
Objective To explore the differentiation of bone marrow mesenchymal stem cells （MSCs） into neuron-llke and dopamlnergic neuron-llke cells after induced by Sonic Hedgehog peptide （Shh）. Methods The MSCs derived from adult human bone marrow were cultured and passaged. After induced with different cytokines and Shh, The number of different immunoreactive cells was detected by nestin, neuron specific enolase （NSE）, glia fibrillary acid protein （GFAP）, tyrosine hydroxylase （TH） and dopamine transporter （DAT） immunocytochemistry. Results Under induction conditions, MSCs resumed typical neuronal morphology. NSE, Nestin,GFAP, TH and DAT were expressed in Neuron-like cells of test group, and were not expressed in MSCs of control group. There were no difference between the percentages of NSE, Nestin and GFAP positive cells in group 1 and that in group 2. The percentages of TH and DAT positive cells in group 2 were higher than that in group 1 （P〈 0.05）. Conclusions After being induced with Shh, the differentiation of human MSCs into dopaminergic neuron-like cells was elevated in vitro.
出处
《中国神经免疫学和神经病学杂志》
CAS
2007年第4期221-223,228,I0004,共5页
Chinese Journal of Neuroimmunology and Neurology
基金
江西省卫生厅科技计划基金资助项目(200501006)
