An adherent-cell depletion technique to generate human neural progenitors and neurons.

An adherent-cell depletion technique to generate human neural progenitors and neurons.
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一种用于产生人类神经祖细胞和神经元的贴壁细胞耗竭技术。

DOI:
10.1002/jcp.28591
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发表时间:
2019
影响因子:
5.6
通讯作者:
Li,JamesYH
Li,JamesYH
中科院分区:
生物学2区
文献类型:
--
作者:
Leung,AlanW;Li,JamesYH

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从胚胎干细胞产生人神经干细胞和神经元的现有方法经常涉及多步骤过程和使用复杂且昂贵的培养基组分、细胞因子或小分子。在这里,我们报告了一种简单的技术,通过定期机械解剖和常规细胞培养级塑料表面上的贴壁细胞去除来产生人神经上皮祖细胞和神经元。这种神经诱导技术不使用生长因子,小分子或肽抑制剂,除了那些存在于无血清补充剂。提示他们的中枢神经系统的起源,我们发现,通过这种技术形成的神经祖细胞表达放射状胶质细胞标记,并在分化时,表达TUBB 3,RBFOX 3(NeuN)和5-羟色胺,但不标记外周神经元。有了这些数据,我们假设周期性机械刺激和塑料表面介导的细胞选择的结合可以改善和简化现有的人类神经元生产方案。
Existing methodologies to produce human neural stem cells and neurons from embryonic stem cells frequently involve multistep processes and the use of complex and expensive media components, cytokines or small molecules. Here, we report a simple technique to generate human neuroepithelial progenitors and neurons by periodic mechanical dissection and adherent‐cell depletion on regular cell‐culture grade plastic surfaces. This neural induction technique does not employ growth factors, small molecules or peptide inhibitors, apart from those present in serum‐free supplements. Suggestive of their central nervous system origin, we found that neural progenitors formed by this technique expressed radial glia markers, and, when differentiated, expressed TUBB3, RBFOX3 (NeuN) and serotonin, but not markers for peripheral neurons. With these data, we postulate that incorporation of periodic mechanical stimuli and plastic surface‐mediated cell selection could improve and streamline existing human neuron production protocols.
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