Lee Minhyung, Sim Hyuna, Ahn Hyunjun, Ha Jeongmin, Baek Aruem, Jeon Young-Joo, Son Mi-Young, Kim Janghwan
Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea.
Department of Functional Genomics, KRIBB, School of Bioscience, University of Science and Technology, Daejeon, Korea.
Int J Stem Cells. 2019 Nov 30;12(3):474-483. doi: 10.15283/ijsc19075.
In Parkinson's disease (PD) research, human neuroblastoma and immortalized neural cell lines have been widely used as models. The advancement in the field of reprogramming technology has provided tools for generating patient-specific induced pluripotent stem cells (hiPSCs) as well as human induced neuronal progenitor cells (hiNPCs). These cells have revolutionized the field of disease modeling, especially in neural diseases. Although the direct reprogramming to hiNPCs has several advantages over differentiation after hiPSC reprogramming, such as the time required and the simple procedure, relatively few studies have utilized hiNPCs. Here, we optimized the protocol for hiNPC reprogramming using pluripotency factors and Sendai virus. In addition, we generated hiNPCs of two healthy donors, a sporadic PD patient, and a familial patient with the G2019S mutation (L2GS). The four hiNPC cell lines are highly proliferative, expressed NPC markers, maintained the normal karyotype, and have the differentiation potential of dopaminergic neurons. Importantly, the patient hiNPCs show different apoptotic marker expression. Thus, these hiNPCs, in addition to hiPSCs, are a favorable option to study PD pathology.
在帕金森病（PD）研究中，人神经母细胞瘤和永生化神经细胞系已被广泛用作模型。重编程技术领域的进展为生成患者特异性诱导多能干细胞（hiPSC）以及人诱导神经祖细胞（hiNPC）提供了工具。这些细胞彻底改变了疾病建模领域，尤其是在神经疾病方面。尽管直接重编程为hiNPC相对于hiPSC重编程后的分化具有几个优点，例如所需时间和操作简单，但利用hiNPC的研究相对较少。在此，我们优化了使用多能性因子和仙台病毒进行hiNPC重编程的方案。此外，我们生成了两名健康供体、一名散发性PD患者和一名携带G2019S突变（L2GS）的家族性患者的hiNPC。这四种hiNPC细胞系具有高度增殖性，表达NPC标志物，维持正常核型，并具有多巴胺能神经元的分化潜能。重要的是，患者hiNPC表现出不同的凋亡标志物表达。因此，这些hiNPC除了hiPSC外，是研究PD病理学的有利选择。
