An interdisciplinary approach and characterization of neuronal cells transdifferentiated from human mesenchymal stem cells
An interdisciplinary approach and characterization of neuronal cells transdifferentiated from human mesenchymal stem cells
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DOI：
10.1089/scd.2007.0011
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发表时间：
2007-10-01
影响因子：
4
通讯作者：
Rameshwar, Pranela
中科院分区：
文献类型：
--
作者：
Greco, Steven J.;Zhou, Chunyi;Rameshwar, Pranela
间充质干细胞（MSCs）是源自中胚层的细胞，主要存在于成人骨髓中。骨髓间充质干细胞表现出谱系分化以产生骨髓间质、脂肪和软骨等细胞。最近的研究报道了间充质干细胞向外胚层和内胚层细胞的转分化。之前，我们已经报道了使用视黄酸（RA）作为分化剂将人(h)间充质干细胞转分化为神经元细胞。本研究提出了一种更有效的诱导方法，并使用分子、细胞和功能方法严格地描述了这种方法的发展。含有碱性成纤维细胞生长因子（bFGF）和RA的诱导剂混合物在诱导后第2天产生表达胶质和神经元祖细胞标志物（GFAP）的细胞。到第12天，90%的hMSCs分化为表达与神经元一致的标记物的细胞，包括与分化神经元发育相关的转录因子。此外，细胞增殖研究和细胞周期特异性蛋白的western blots表明，第12天诱导的细胞是有丝分裂后细胞，没有细胞死亡的证据。这些细胞表现出自发的突触后电流，能够合成、包装和释放神经递质。总之，改进的诱导方案，结合跨学科的方法来验证hMSCs可以分化为神经细胞，为再生医学模型的转化应用提供了一步。
Mesenchymal stem cells (MSCs) are mesoderm-derived cells, primarily resident in adult bone marrow (BM). MSCs exhibit lineage differentiation to generate cells such as BM stroma, fat, and cartilage. Recent studies have reported the transdifferentiation of MSCs to cells of ectodermal and endodermal origin. Previously, we have reported transdifferentiation of human (h) MSCs into neuronal cells using retinoic acid (RA) as a differentiating agent. This study presents a more efficient induction method and rigorously characterizes the development using molecular, cellular, and functional approaches. A cocktail of induction agents containing basic fibroblast growth factor (bFGF) and RA generated cells that expressed glial and neuronal progenitor markers (GFAP) at day 2 post-induction. By day 12, 90% of hMSCs differentiated into cells that expressed markers consistent for neurons, including transcription factors linked to the development of differentiated neurons. Furthermore, cell proliferation studies and western blots for cell cycle-specific proteins demonstrated day-12 induced cells to be post-mitotic cells with no evidence of cell death. The cells exhibited spontaneous post-synaptic currents and were capable of neurotransmitter synthesis, packaging, and release. Together, the improved induction protocol, combined with an interdisciplinary approach to verify that hMSCs can differentiate into neuronal cells, provides a step toward translational application with models of regenerative medicine.