Differentiation of dopaminergic neurons from human embryonic stem cells: Modulation of differentiation by FGF-20
Differentiation of dopaminergic neurons from human embryonic stem cells: Modulation of differentiation by FGF-20
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DOI：
10.1016/j.jbiosc.2008.12.013
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发表时间：
2009-04-01
影响因子：
2.8
通讯作者：
Kunisada, Takahiro
中科院分区：
文献类型：
--
作者：
Shimada, Hideaki;Yoshimura, Naoko;Kunisada, Takahiro
由于中脑多巴胺能（DA）神经元移植治疗帕金森病（PD）的临床潜力，中脑多巴胺能（DA）神经元从人胚胎干（hES）细胞中衍生一直受到特别关注。已经报道了几种从小鼠胚胎干细胞和he细胞分化DA神经元的方案，然而，涉及he细胞的方案尚未得到改进。在这里，我们使用稍微改进的基质细胞衍生的诱导活性方法，包括四个不同的培养阶段，以证明KhES-1细胞分化为酪氨酸羟化酶（TH）阳性的DA神经元。实时荧光定量PCR分析显示，DA神经元标记基因NURR1、配对样同源结构域转录因子3 （PITX3）、LIM同源盒转录因子1、β (LMX1B)、EN1 （EN1）、多巴胺转运蛋白（DAT）和芳香氨基酸脱羧酶（AADC）在分化过程中被显著诱导。在最终分化阶段用成纤维细胞生长因子(FGF)-20和FGF-2处理可诱导DA神经元发育相关转录因子如NURR1、PITX3、LMX1B和EN1的增加。FGF-20和FGF-2直接促进了来自hES细胞的神经祖细胞的DA神经元分化，而不需要任何来自PA6细胞的可溶性因子。这些结果提供了有价值的信息，将有助于从he细胞高效DA神经元分化和未来的移植应用。(c) 2009，生物技术学会，日本。版权所有。
Derivation of midbrain dopaminergic (DA) neurons from human embryonic stem (hES) cells has been of particular interest because of the clinical potential for DA neuron transplantation in patients with Parkinson's disease (PD). Several protocols for DA neuron differentiation from mouse embryonic stem cells and hES cells have been reported: however, protocols involving hES cells have yet to be improved. Here, we used a slightly modified stromal cell-derived inducing activity method, consisting four different culture stages, to show that KhES-1 cells differentiate into tyrosine hydroxylase (TH)-positive DA neurons. Quantitative real-time PCR analysis showed a marked induction of the DA neuron marker genes NURR1, paired-like homeodomain transcription factor 3 (PITX3), LIM homeobox transcription- factor 1, beta (LMX1B), engrailed-1 (EN1), dopamine transporter (DAT, and aromatic amino acid decarboxylase (AADC) during differentiation. Treatment with fibroblast growth factor (FGF)-20 and FGF-2 at the final differentiation stage induced the increase of DA neuron development-related transcription factors such as NURR1, PITX3, LMX1B, and EN1. FGF-20 and FGF-2 enhanced DA neuron differentiation from hES cell-derived neural progenitor cells directly without any soluble factors from PA6 cells. These results provide valuable information that will assist in efficient DA neuron differentiation from hES cells and for future transplant application. (c) 2009, The Society for Biotechnology, Japan. All rights reserved.