Plasticity of cultured mesenchymal stem cells: Switch from nestin-positive to excitable neuron-like phenotype
Plasticity of cultured mesenchymal stem cells: Switch from nestin-positive to excitable neuron-like phenotype
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DOI：
10.1634/stemcells.2004-0149
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发表时间：
2005-03-01
期刊：
影响因子：
5.2
通讯作者：
Rogister, B
中科院分区：
文献类型：
--
作者：
Wislet-Gendebien, S;Hans, G;Rogister, B
骨髓间充质干细胞（MSCs）可以分化为多种类型的间充质细胞，包括骨细胞，软骨细胞和脂肪细胞，但在适当的实验条件下，也可以分化为非间充质细胞，如神经细胞。这些观察结果引起了人们对MSC在各种神经系统疾病的细胞治疗策略中的可能用途的兴趣。在这里报道的研究中，我们解决了MSC体外分化为功能性神经元的问题。首先，我们证明，当它们与小脑颗粒神经元共培养时，成人MSC可以表达神经元标记物。MSCs向神经元分化需要两个因素：第一个因素是MSCs表达巢蛋白（巢蛋白是MSCs对外部信号的反应特征的标志物），第二个因素是神经细胞和MSCs之间的直接细胞-细胞相互作用，允许这些外部信号的整合。三种不同的方法表明，神经表型是由MSC分化而不是细胞融合过程产生的，尽管最后一种现象也可以共存。通过定量逆转录聚合酶链反应（RT-PCR）分析了几个基因（包括sox、pax、notch、delta、frizzled和erbB）的表达，以进一步表征nestin阳性表型与nestin阴性表型相比。在nestin阳性的MSC中发现sox 2、sox 10、pax 6、fzd、erbB 2和erbB 4的过表达。最后，电生理分析表明，MSC衍生的神经元样细胞可以激发单动作电位并对GABA、甘氨酸和谷氨酸等多种神经递质做出反应。我们的结论是，巢蛋白阳性的骨髓间充质干细胞可以在体外分化为可兴奋的神经元样细胞。
Bone marrow mesenchymal stem cells (MSCs) can differentiate into several types of mesenchymal cells, including osteocytes, chondrocytes, and adipocytes, but, under appropriate experimental conditions, can also differentiate into nonmesenchymal cells-for instance, neural cells. These observations have raised interest in the possible use of MSCs in cell therapy strategies for various neurological disorders. In the study reported here, we addressed the question of in vitro differentiation of MSCs into functional neurons. First, we demonstrate that when they are co-cultured with cerebellar granule neurons, adult MSCs can express neuronal markers. Two factors are needed for the emergence of neuronal differentiation of the MSCs: the first one is nestin expression by MSCs (nestin is a marker for the responsive character of MSCs to extrinsic signals), and the second one is a direct cell-cell interaction between neural cells and MSCs that allows the integration of these extrinsic signals. Three different approaches suggest that neural phenotypes arise from MSCs by a differentiation rather than a cell fusion process, although this last phenomenon can also coexist. The expression of several genes-including sox, pax, notch, delta,frizzled, and erbB-was analyzed by quantitative reverse transcription polymerase chain reaction (RT-PCR) in order to further characterize the nestin-positive phenotype compared to the nestin-negative one. An overexpression of sox2, sox10, pax6, fzd, erbB2, and erbB4 is found in nestin-positive MSCs. Finally, electrophysiological analyses demonstrate that MSC-derived neuron-like cells can fire single-action potentials and respond to several neurotransmitters such as GABA, glycine, and glutamate. We conclude that nestin-positive MSCs can differentiate in vitro into excitable neuron-like cells.