Rapid Conversion of Human Induced Pluripotent Stem Cells into Dopaminergic Neurons by Inducible Expression of Two Transcription Factors
Rapid Conversion of Human Induced Pluripotent Stem Cells into Dopaminergic Neurons by Inducible Expression of Two Transcription Factors
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DOI：
10.1089/scd.2021.0363
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发表时间：
2022-05-12
影响因子：
4
通讯作者：
Takata, Kazuyuki
中科院分区：
文献类型：
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作者：
Nishimura, Kaneyasu;Nitta, Tatsumi;Takata, Kazuyuki
人多能干细胞（hPSC），包括人胚胎干细胞和人诱导多能干细胞（hiPSC），为再生治疗、疾病建模和药物筛选提供了有希望的来源。相关的努力已经投入到通过模拟大脑发育相关的信号通路来建立稳健的诱导方案，用于PSC衍生的多巴胺（DA）神经元的产生。然而，这些方案需要经过充分训练的技术和很长的时间来产生成熟的DA神经元。在这项研究中，为了加速整个过程，我们通过两种转录因子ASCL1和LMX1A的诱导力表达产生了分化为DA神经元的hiPSC系。使用该hiPSC系，我们建立了快速且简单的诱导方案以在28天内产生成熟的DA神经元。诱导的DA神经元的特点是基因表达和免疫组化分析的基本DA神经元标志物。此外，在第28天通过多电极阵列系统分析细胞功能特性。该资源为高通量筛选提供了未来的应用，例如需要高度验证的DA神经元的药物开发和毒理学。
Human pluripotent stem cells (hPSCs), including human embryonic stem cells and human induced pluripotent stem cells (hiPSCs), provide promising sources for regenerative therapy, disease modeling, and drug screening. Relevant efforts have been invested in establishing robust induction protocols for PSC-derived dopaminergic (DA) neuron generation by mimicking brain development-related signaling pathways. However, these protocols require fully trained techniques and a long time to yield mature DA neurons. In this study, to accelerate the entire process, we generated a hiPSC line differentiating into DA neurons by the inducible force expression of two transcription factors ASCL1 and LMX1A. Using this hiPSC line, we established a rapid and simple induction protocol to generate mature DA neurons in 28 days. The induced DA neurons were characterized by gene expression and immunohistochemical analyses of fundamental DA neuronal markers. Moreover, the cell functional properties were analyzed by a multielectrode array system on day 28. This resource offers future applications for high-throughput screening, such as drug development and toxicology that require highly validated DA neurons.