New methods for inducing the differentiation of amniotic-derived mesenchymal stem cells into motor neuron precursor cells
New methods for inducing the differentiation of amniotic-derived mesenchymal stem cells into motor neuron precursor cells
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诱导羊膜间充质干细胞分化为运动神经元前体细胞的新方法
DOI：
10.1016/j.tice.2013.03.002
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发表时间：
2013-10-01
期刊：
影响因子：
2.6
通讯作者：
Yang Bo
中科院分区：
文献类型：
--
作者：
Hu Wei;Guan Fang-xia;Yang Bo
本研究探讨了细胞外基质（extracellular matrix，ECM）和多种细胞因子联合诱导骨髓间充质干细胞（amenchymal stem cells，ADMSCs）向运动神经元（motor neuron，MN）前体细胞分化的作用。采用酶法和化学法制备了具有良好生物相容性的膜状细胞外基质。实验组（EG）中的细胞用ECM和多细胞因子在多步诱导过程中处理，而对照组（CG）的处理类似，但不使用ECM。在EG中，诱导后，细胞形成突起，与相邻细胞连接，形成具有方向性的网络。在这些细胞中，神经元特异性烯醇化酶（NSE）和突触素（SYN）的表达水平增加，胶质细胞酸性蛋白（GFAP）的表达减少。EG细胞中SYN的表达高于CG细胞。在对照组中，NSE表达增加，而Nestin和SYN的表达没有变化。这些是其他基因水平的几个变化：第1代ADMSCs表达Nanog、SOX 2、八聚体结合转录因子4（OCT 4）和Nestin。在EG中，在诱导开始时，Nanog的表达降低，SOX 2和Nestin的表达增加。2天后，细胞表达Nestin、OCT 4和SYNIII，3天后，它们表达Olig 2、OCT 4、Nestin、SYNII和Islet 1（ISL 1）。最后，在第6天，细胞表达巢蛋白、SYNI、SYNIII、ISL-1、同源框9（Hb 9）和少突胶质细胞谱系转录因子2（Olig 2）。在CG中，细胞从不表达SYNI、SYNII或Hb 9。因此，我们的研究表明，提取的ECM能够促进突触的成熟。人ADMSC由多种细胞亚群组成，包括神经祖细胞。本研究中使用的多步诱导方法使人ADMSC分化为MN前体细胞。皇冠版权所有（C）2013由爱思唯尔有限公司出版。保留所有权利。
Our study investigates the differentiation of amniotic-derived mesenchymal stem cells (ADMSCs) into motor neuron (MN) precursor cells induced by a combination of extracellular matrix (ECM) and multi-cell factors. Membrane-like ECM was made by an enzymatic and chemical extraction method and exhibited good biological compatibility. Cells in the experimental group (EG) were treated with ECM and multi-cell factors in a multi-step induction process, while the control group (CG) was treated similarly, except without ECM. In the EG, after induction, the cells formed processes that connected with neighboring cells to form a net that had directionality. In these cells, neuron-specific enolase (NSE) and synaptophysin (SYN) expression levels increased and glial fibrillary acidic protein (GFAP) expression decreased. The SYN expression in the EG cells was higher compared with those in the CG. In the CG, NSE expression increased, while the expression of Nestin and SYN did not change. These were several changes in the levels of other genes: ADMSCs at passage 1 expressed Nanog, SOX2, octamer-binding transcription factor 4 (OCT4) and Nestin. In the EG, at the beginning of induction, the expression of Nanog decreased and that of SOX2 and Nestin increased. After 2 days, the cells expressed Nestin, OCT4 and SYNIII, and after 3 days, they expressed Olig2, OCT4, Nestin, SYNII and Islet1 (ISL1). Finally, at day 6, the cells expressed Nestin, SYNI, SYNIII, ISL-1, homeobox 9 (Hb9) and oligodendrocyte lineage transcription factor 2 (Olig2). In the CG, the cells never expressed SYNI, SYNII or Hb9. Our studies therefore demonstrate that the extracted ECM was capable of promoting the maturation of synapses. Human ADMSCs are composed of multiple cell subsets, including neural progenitor cells. The multi-step induction method used in this study causes human ADMSCs to differentiate into MN precursor cells. Crown Copyright (C) 2013 Published by Elsevier Ltd. All rights reserved.