Institute of Molecular and Cell Biology, A*STAR Research Entities, Singapore, Singapore.
NUS Graduate School for Integrative Science and Engineering, National University of Singapore, Singapore, Singapore.
Methods Mol Biol. 2022;2549:359-377. doi: 10.1007/7651_2021_399.
The use of patient-derived induced pluripotent stem cells (iPSCs) and their neural derivatives is becoming increasingly important in the study of neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, Lewy body dementia, amyotrophic lateral sclerosis, peripheral neuropathy, and so on. Increasingly, iPSC-derived neurons also reveal key pathways and signaling defects in psychiatric disorders such as autism spectrum disorders, schizophrenia, and bipolar disorder. With recent advances in CRISPR/Cas9-mediated genome editing technology, patient-derived iPSCs with disease-causing mutations can be corrected into "isogenic control lines," and these can be differentiated into neural derivatives with identical genetic background. This provides an opportunity for in vitro disease modeling to unravel disease mechanisms and a platform to facilitate drug discovery. In this chapter, we provide details of the differentiation protocols to reliably derive four currently relevant neuronal subtypes, i.e., cortical neurons, midbrain dopaminergic neurons, spinal motor neurons, and sensory neurons.
使用患者来源的诱导多能干细胞（iPSC）及其神经衍生物在研究神经退行性疾病（如阿尔茨海默病、帕金森病、路易体痴呆、肌萎缩侧索硬化症、周围神经病等）方面变得越来越重要。越来越多的 iPSC 衍生神经元也揭示了精神疾病（如自闭症谱系障碍、精神分裂症和双相情感障碍）中的关键途径和信号缺陷。随着 CRISPR/Cas9 介导的基因组编辑技术的最新进展，具有致病突变的患者来源 iPSC 可以被纠正为“同基因对照系”，并可以将其分化为具有相同遗传背景的神经衍生物。这为体外疾病建模提供了机会，以揭示疾病机制，并为药物发现提供了平台。在本章中，我们提供了详细的分化方案，以可靠地获得目前相关的四种神经元亚型，即皮质神经元、中脑多巴胺能神经元、脊髓运动神经元和感觉神经元。
