Induction of GABAergic phenotype in a neural stem cell line for transplantation in an excitotoxic model of Huntington's disease
Induction of GABAergic phenotype in a neural stem cell line for transplantation in an excitotoxic model of Huntington's disease
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DOI：
10.1016/j.expneurol.2004.06.027
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发表时间：
2004-11-01
影响因子：
5.3
通讯作者：
Canals, JM
中科院分区：
文献类型：
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作者：
Bosch, M;Pineda, JR;Canals, JM
使用多能神经干细胞(NSCs)实施细胞替代疗法治疗亨廷顿病需要在移植前特定分化为伽马氨基丁酸(GABA)神经元亚型。在这里，我们提出了一种有效的培养方法，可以从永生化的纹状体神经干细胞系ST14A中诱导出稳定的GABA能神经元。这一过程需要连续的维甲酸处理和氯化钾去极化。最初加入10um维甲酸可增加细胞存活率并促进神经元分化。随后用40 mM KCl刺激诱导分化为GABA能神经元，占培养细胞总数的74%。KCL诱导的钙内流抑制细胞增殖和巢蛋白表达，诱导轴突生长和GABA能标志物以及GABA含量、释放和摄取。在亨廷顿病模型中，将这些预分化的GABA能神经元移植到成年脑中后，对其整合、存活和表型的特征表明，在喹啉损伤的纹状体内可以长期存活。在这些条件下，细胞维持其GABA能表型，并通过与内源性神经元的突触接触来阐述突起过程。总而言之，我们已经从神经干细胞系中产生了一个同质的功能GABA能神经元群体，这些神经元在体内存活并维持其获得性命运。这些数据可能支持使用神经干细胞对亨廷顿病进行细胞替代疗法的可能性。(C)2004 Elsevier Inc.保留所有权利。
The implementation of cell replacement therapies for Huntington's disease using multipotent neural stem cells (NSCs) requires the specific differentiation into gamma-aminobutyric acid (GABA) neuronal subtype before transplantation. Here we present an efficient culture procedure that induces stable GABAergic neurons from the immortalized striatal neural stem cell line ST14A. This process requires sequential retinoic acid treatment and KCl depolarization. Initial addition of 10 muM retinoic acid increased cell survival and promoted neuronal differentiation. Subsequent stimulation with 40 mM KCl induced specific differentiation into GABAergic neurons, yielding 74% of total cultured cells. KCl-evoked Ca2+ influx reduced cell proliferation and nestin expression, and induced neurite outgrowth and GABAergic markers as well as GABA contents, release, and uptake. Characterization of the integration, survival, and phenotype of these predifferentiated GABAergic neurons following transplantation into the adult brain in a model of Huntington's disease revealed long-term survival in quinolinate-lesioned striata. Under these conditions, cells maintained their GABAergic phenotype and elaborated neurite processes with synaptic contacts with endogenous neurons. In conclusion, we have generated a homogeneous population of functional GABAergic neurons from a neural stem cell line, which survive and maintain their acquired fate in vivo. These data may lend support to the possibility of cell replacement therapies for Huntington's disease using neural stem cells. (C) 2004 Elsevier Inc. All rights reserved.