Generation of neurons from human induced pluripotent stem cells (hiPSCs) overcomes the limited access to human brain samples and greatly facilitates the progress of research in neurological diseases. However, it is still a challenge to generate a particular neuronal subtype with high purity and yield for determining the pathogenesis of diseased neurons using biochemical approaches. Motor neurons (MNs) are a specialized neuronal subtype responsible for governing both autonomic and volitional movement. Dysfunctions in MNs are implicated in a variety of movement diseases, such as amyotrophic lateral sclerosis (ALS). In this study, we generated functional MNs from human iPSCs via lentiviral delivery of transcription factors. Moreover, we optimized induction conditions by using different combinations of transcription factors and found that a single lentiviral vector expressing three factors (NGN2, ISL1, and LHX3) is necessary and sufficient to induce iPSC-MNs. These MNs robustly expressed general neuron markers (MAP2, SMI-32, and TUBB3), MN-specific markers (HB9 and ChAT), and showed electrical maturation and firing of action potentials within 3 weeks. This approach significantly improved the neuronal survival, yield, and purity, making it feasible to obtain abundant materials for biochemical studies in modeling movement diseases.

中文翻译：

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通过慢病毒传递转录因子从人诱导多能干细胞中生成和优化高纯度运动神经元。


人类诱导多能干细胞（hiPSC）产生神经元克服了人类大脑样本获取的有限性，极大地促进了神经系统疾病研究的进展。然而，产生高纯度和高产量的特定神经元亚型以利用生化方法确定患病神经元的发病机制仍然是一个挑战。运动神经元（MN）是一种特殊的神经元亚型，负责控制自主运动和意志运动。 MN 的功能障碍与多种运动疾病有关，例如肌萎缩侧索硬化症 (ALS)。在这项研究中，我们通过慢病毒传递转录因子从人类 iPSC 中产生了功能性 MN。此外，我们通过使用不同的转录因子组合优化诱导条件，发现表达三个因子（NGN2、ISL1和LHX3）的单个慢病毒载体对于诱导iPSC-MN是必要且充分的。这些 MN 强烈表达一般神经元标记（MAP2、SMI-32 和 TUBB3）、MN 特异性标记（HB9 和 ChAT），并在 3 周内表现出电成熟和动作电位放电。这种方法显着提高了神经元的存活率、产量和纯度，使得获得丰富的材料用于运动疾病建模的生化研究成为可能。