Accelerated high-yield generation of limb-innervating motor neurons from human stem cells.
Accelerated high-yield generation of limb-innervating motor neurons from human stem cells.
复制标题
DOI：
10.1523/jneurosci.0906-12.2013
复制
发表时间：
2013-01-09
期刊：
影响因子：
--
通讯作者：
Wichterle H
中科院分区：
文献类型：
--
作者：
Amoroso MW;Croft GF;Williams DJ;O'Keeffe S;Carrasco MA;Davis AR;Roybon L;Oakley DH;Maniatis T;Henderson CE;Wichterle H
人类多能干细胞是发育研究、细胞移植、疾病建模和药物测试中有前途的分化细胞来源。然而，即使对脊髓运动神经元等已深入研究的细胞类型，它们的广泛应用也受到现有体外分化方案持续时间长、产量低以及所产生的群体分子异质性的阻碍。我们报告了一种小分子组合，在3周内诱导运动神经元丰度高达50%，并具有与神经退行性疾病相关的明确亚型身份。尽管它们的分化加速，但运动神经元表达HB9、ISL1和柱特异性标记的组合，这与在人胎儿脊髓体内观察到的情况相一致。它们也表现出自发和诱导的活动，并在移植到发育中的小鸡脊髓时向肌肉投射轴突。引人注目的是，这种新方案优先产生表达肢体支配外侧运动柱运动神经元标记物（FOXP1+/LHX3−）的运动神经元。获得人类肢体支配运动神经元亚型的高产培养将促进运动神经元亚型特异性特性的深入研究、疾病建模和大规模基于细胞的筛选分析的发展。
Human pluripotent stem cells are a promising source of differentiated cells for developmental studies, cell transplantation, disease modeling, and drug testing. However, their widespread use even for intensely studied cell types like spinal motor neurons is hindered by the long duration and low yields of existing protocols for in vitro differentiation and by the molecular heterogeneity of the populations generated. We report a combination of small molecules that within 3 weeks induce motor neurons at up to 50% abundance and with defined subtype identities of relevance to neurodegenerative disease. Despite their accelerated differentiation, motor neurons expressed combinations of HB9, ISL1 and column-specific markers that mirror those observed in vivo in human fetal spinal cord. They also exhibited spontaneous and induced activity, and projected axons towards muscles when grafted into developing chick spinal cord. Strikingly, this novel protocol preferentially generates motor neurons expressing markers of limb-innervating lateral motor column motor neurons (FOXP1+/LHX3−). Access to high-yield cultures of human limb-innervating motor neuron subtypes will facilitate in-depth study of motor neuron subtype-specific properties, disease modeling, and development of large-scale cell-based screening assays.