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首页> 外文期刊>Journal of biomedical science. >Fibroblast Growth Factor-2 alone as an efficient inducer for differentiation of human bone marrow mesenchymal stem cells into dopaminergic neurons
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Fibroblast Growth Factor-2 alone as an efficient inducer for differentiation of human bone marrow mesenchymal stem cells into dopaminergic neurons

机译：单独的成纤维细胞生长因子2是有效的诱导人骨髓间充质干细胞分化为多巴胺能神经元的诱导剂

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BackgroundThe reported efficiency of differentiation of human bone marrow derived Mesenchymal Stem Cells (hBM MSC) into dopaminergic neurons with different inducers is found to vary. Thus, in the current study we have investigated the response of hBM MSC to some of the neuronal inducers and their combinations. Neuronal differentiation inducing agents Fibroblastic Growth Factor 2 (FGF2), Sonic Hedge Hog (Shh), Fibroblastic Growth Factor 8 (FGF8) & All Trans Retinoic Acid (ATRA) were used either singly or in varied combinations.ResultsThe differentiated and undifferentiated hBM MSC were characterized in terms of morphology, expression of cell markers at transcriptional and translational levels, amount of dopamine secreted by the cells in the media and changes in cell membrane potential by calcium ions imaging. Induced hBM MSC revealed neuron like morphology and expressed cellular markers suggesting neuronal differentiation with all the inducing agents. However, upon quantitative analysis through qPCR, cells induced with FGF2 were found to show maximum expression of tyrosine hydroxylase (TH) by 47.5 folds. Immunofluorescence analysis of differentiated and undifferentiated cells also revealed expression of nestin, neurofilament, microtubule associated protein- 2, beta tubulin III and TH in differentiated cells, at translational level. This data was supported by immunoblotting analysis. Further, ELISA study also supported the release of dopamine by cultures induced with FGF2. When the cells were depolarised with KCl solution, those induced with Shh & FGF8 showed maximum calcium ion trafficking, followed by the cells induced with FGF2 only.ConclusionsWe conclude that hBM MSC can be coaxed to differentiate efficiently into dopaminergic neurons in the presence of a very simple media cocktail containing only one main inducer like FGF2 and thus contribute towards cellular therapy in Parkinson's and other related disorders. These dopaminergic neurons are also functionally active, as shown by calcium ion trafficking.Electronic supplementary materialThe online version of this article (doi:10.1186/s12929-014-0083-1) contains supplementary material, which is available to authorized users.
机译：背景技术据报道，人类骨髓间充质干细胞（hBM MSC）分化为具有不同诱导物的多巴胺能神经元的效率有所不同。因此，在当前的研究中，我们研究了hBM MSC对某些神经元诱导剂及其组合的反应。神经分化诱导剂成纤维细胞生长因子2（FGF2），Sonic Hedge Hog（Shh），成纤维细胞生长因子8（FGF8）和全反式维甲酸（ATRA）单独使用或以多种组合使用。结果分化和未分化的hBM MSC分别为根据形态，细胞标志物在转录和翻译水平的表达，培养基中细胞分泌的多巴胺的量以及钙离子成像显示的细胞膜电位的变化进行特征分析。诱导的hBM MSC揭示了神经元样形态，并表达了细胞标志物，暗示了所有诱导剂的神经元分化。但是，通过qPCR进行定量分析后，发现FGF2诱导的细胞显示酪氨酸羟化酶（TH）的最大表达达47.5倍。对分化和未分化细胞的免疫荧光分析还揭示了在翻译水平上巢蛋白，神经丝，微管相关蛋白2，β微管蛋白III和TH在分化细胞中的表达。该数据得到了免疫印迹分析的支持。此外，ELISA研究还支持FGF2诱导的培养物释放多巴胺。当用KCl溶液使细胞去极化时，用Shh和FGF8诱导的细胞显示出最大的钙离子运输，其次是仅用FGF2诱导的细胞。简单的培养基混合物，仅包含一种主要的诱导物，如FGF2，因此有助于帕金森氏症和其他相关疾病的细胞治疗。这些多巴胺能神经元也具有功能活性，如钙离子贩运所示。电子补充材料本文的在线版本（doi：10.1186 / s12929-014-0083-1）包含补充材料，授权用户可以使用。

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