Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
Cells. 2022 Nov 27;11(23):3796. doi: 10.3390/cells11233796.
Generation of motor neurons (MNs) from human-induced pluripotent stem cells (hiPSCs) overcomes the limited access to human brain tissues and provides an unprecedent approach for modeling MN-related diseases. In this review, we discuss the recent progression in understanding the regulatory mechanisms of MN differentiation and their applications in the generation of MNs from hiPSCs, with a particular focus on two approaches: induction by small molecules and induction by lentiviral delivery of transcription factors. At each induction stage, different culture media and supplements, typical growth conditions and cellular morphology, and specific markers for validation of cell identity and quality control are specifically discussed. Both approaches can generate functional MNs. Currently, the major challenges in modeling neurological diseases using iPSC-derived neurons are: obtaining neurons with high purity and yield; long-term neuron culture to reach full maturation; and how to culture neurons more physiologically to maximize relevance to in vivo conditions.
从人诱导多能干细胞 (hiPSC) 中生成运动神经元 (MN) 克服了人类脑组织获取的限制，并为 MN 相关疾病的建模提供了一种前所未有的方法。在这篇综述中，我们讨论了理解 MN 分化调控机制的最新进展及其在 hiPSC 中生成 MN 的应用，特别关注两种方法：小分子诱导和慢病毒转导转录因子诱导。在每个诱导阶段，我们详细讨论了不同的培养基和补充剂、典型的生长条件和细胞形态，以及用于验证细胞身份和质量控制的特异性标记物。这两种方法都可以生成功能性 MN。目前，使用 iPSC 衍生神经元进行神经疾病建模的主要挑战是：获得高纯度和高产量的神经元；长期神经元培养以达到完全成熟；以及如何更生理地培养神经元，以最大限度地提高与体内条件的相关性。
