隐丹参酮诱导猴骨髓间质干细胞分化为神经元样细胞

Macaca irus bone mesenchymal stem cells differentiate into neuron-like cells induced by cryptotanshinone in vitro

目的体外培养猴骨髓间质干细胞(MSCs)并定向诱导分化为神经元样细胞。方法体外分离培养猴MSCs,流式细胞仪检测其表面标志,用含隐丹参酮的无血清L-DMEM诱导MSCs分化为神经元样细胞,免疫细胞化学检测单克隆抗体特异性神经元烯醇化酶(NSE)、神经丝蛋白(NF)、胶质纤维酸性蛋白(GFAP)的表达。结果体外培养的猴MSCs表达CD29、CD44、CD105、CD166,并且具有正常的二倍体核型,不随传代而发生改变。bFGF预诱导24h后,隐丹参酮可以将MSCs诱导为神经元样细胞,免疫细胞化学显示NSE、NF表达阳性,阳性率分别为68.3%±3.5%、70.3%±1.5%,而GFAP表达阴性。结论隐丹参酮可以在体外将猴MSCs诱导分化为神经元样细胞。

AIM: To determine the optimal protocol and condition in which macaca irus mesenchymal stem cells (MSCs) are induced to differentiate into neuron-like cells by cryptotanshinone in vitro. METHODS: MSCs from macaca irus bone marrow were generated in vitro and induced with cryptotanshinone. The morphological changes of MSCs were evaluated by microscope. The positive percentages of neurofilament (NF), neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression were measured by immunocytochemistry with ABC staining. RESULTS: The result showed that MSCs were positive for CD29, CD44, CD105, CD166, and negative for CD34, CD71, CD80 and CD86. After induced with cryptotanshinone, MSCs began to display neuronal morphologies, such as contracted multipolar cell body and formed extensive networks. The percentages of positive NSE, NF expression were 68.3%±3.5%, 70.3%±1.5%, respectively. CONCLUSION: Macaca irus MSCs could be induced to differentiate into neuron-like cells in vitro by cryptotanshinone and might be applied in cell transplantation and gene therapy in nervous system disorders.