Stem Cells Expanded from the Human Embryonic Hindbrain Stably Retain Regional Specification and High Neurogenic Potency
Stem Cells Expanded from the Human Embryonic Hindbrain Stably Retain Regional Specification and High Neurogenic Potency
复制标题
DOI：
10.1523/jneurosci.0130-13.2013
复制
发表时间：
2013-07-24
影响因子：
5.3
通讯作者：
Smith, Austin
中科院分区：
文献类型：
--
作者：
Tailor, Jignesh;Kittappa, Raja;Smith, Austin
干细胞系忠实地维持人类大脑中前体细胞的区域特性和发育潜力，将在发育神经生物学领域创造新的机会，并为生成专门的人类神经元提供资源。然而，到目前为止，来源于人脑的神经前体细胞培养要么寿命很短，要么表现出有限的分化能力，主要是神经胶质细胞。多能干细胞是另一种来源，但要确定仅在体外产生的细胞类型的身份和保真度是有问题的。在这里，我们展示了后脑神经上皮干细胞(HbNES)可以从早期的人类胚胎(5-7周，卡内基阶段15-17)获得并大量扩增。这些细胞系在EGF和FGF2存在的贴壁培养中增殖，并保留了祖细胞特征，包括SOX1表达，形成玫瑰花样结构，以及高神经分化能力。它们产生GABA能、谷氨酸能神经元，频率较低时产生5-羟色胺能神经元。重要的是，hbNES细胞稳定地维持后脑的特性，并在暴露于骨形态发生蛋白(BMP)的情况下产生上菱形嘴唇衍生物。当它们被移植到新生大鼠的大脑中时，它们显示出整合到小脑发育中的潜力，并产生小脑粒样细胞，尽管频率很低。HbNES细胞为研究人类小脑的规范和发育以及模拟后脑疾病提供了一个新的系统。它们还为从多能细胞系生产类似的长期神经上皮样干细胞(lt-NES)提供了基准。据我们所知，hbNES细胞是从人类胚胎中获得高度可扩展的神经上皮干细胞的第一个证据，而不是遗传永生化。
Stem cell lines that faithfully maintain the regional identity and developmental potency of progenitors in the human brain would create new opportunities in developmental neurobiology and provide a resource for generating specialized human neurons. However, to date, neural progenitor cultures derived from the human brain have either been short-lived or exhibit restricted, predominantly glial, differentiation capacity. Pluripotent stem cells are an alternative source, but to ascertain definitively the identity and fidelity of cell types generated solely in vitro is problematic. Here, we show that hindbrain neuroepithelial stem (hbNES) cells can be derived and massively expanded from early human embryos (week 5-7, Carnegie stage 15-17). These cell lines are propagated in adherent culture in the presence of EGF and FGF2 and retain progenitor characteristics, including SOX1 expression, formation of rosette-like structures, and high neurogenic capacity. They generate GABAergic, glutamatergic and, at lower frequency, serotonergic neurons. Importantly, hbNES cells stably maintain hindbrain specification and generate upper rhombic lip derivatives on exposure to bone morphogenetic protein (BMP). When grafted into neonatal rat brain, they show potential for integration into cerebellar development and produce cerebellar granule-like cells, albeit at low frequency. hbNES cells offer a new system to study human cerebellar specification and development and to model diseases of the hindbrain. They also provide a benchmark for the production of similar long-term neuroepithelial-like stem cells (lt-NES) from pluripotent cell lines. To our knowledge, hbNES cells are the first demonstration of highly expandable neuroepithelial stem cells derived from the human embryo without genetic immortalization.