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Epidermal growth factor-immobilized surfaces for the selective expansion of neural progenitor cells derived from induced pluripotent stem cells.
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2020-05-30 , DOI: 10.1002/bit.27445
Yuka Yamauchi 1, 2 , Isao Hirata 1 , Kotaro Tanimoto 2 , Koichi Kato 1
Affiliation  

Neural progenitor cells (NPCs) are considered to be a promising source for stem cell‐based regenerative therapy for central nervous disorders. However, the widespread clinical application of NPCs requires another technology that permits the efficient production of pure NPCs in large quantities. In this study, culture substrates were designed by immobilizing epidermal growth factor (EGF) onto the substrate and evaluated for their feasibility of expanding NPCs obtained through the neurosphere culture of induced pluripotent stem (iPS) cells. After three passages we obtained neurospheres that contained cells abundantly expressing an EGF receptor. The neurospheres were dissociated into single cells and seeded onto the EGF‐immobilized substrates. It was observed that neurosphere‐forming cells seeded and cultured on the EGF‐immobilized surface formed a two‐dimensional cellular network characteristic of NPCs. These cells were found to be capable of being subcultured, while remaining their proliferation potential. Furthermore, a majority of cells (~99% of total cells) on the substrate was shown to express an NPC marker, nestin, whereas a limited number of cells (~1% of total cells) expressed neuronal marker, β‐tubulin III. These results as a whole demonstrate that the EGF‐immobilized substrate allows for iPS cell‐derived NPCs to efficiently proliferate while maintaining the undifferentiated state.

中文翻译:

表皮生长因子固定化表面,用于选择性扩增源自诱导多能干细胞的神经祖细胞。

神经祖细胞(NPC)被认为是基于干细胞的中枢神经疾病再生疗法的有希望的来源。然而,NPCs 的广泛临床应用需要另一种技术来允许大量高效生产纯 NPCs。在这项研究中,通过将表皮生长因子 (EGF) 固定在基质上来设计培养基质,并评估它们通过诱导多能干 (iPS) 细胞的神经球培养获得的扩大 NPC 的可行性。三代后,我们获得了含有大量表达 EGF 受体的细胞的神经球。神经球被分解成单个细胞并接种到 EGF 固定的基质上。观察到在 EGF 固定化表面接种和培养的神经球形成细胞形成了 NPC 的二维细胞网络特征。发现这些细胞能够进行传代培养,同时保持其增殖潜力。此外,基底上的大多数细胞(约占总细胞的 99%)表达 NPC 标记物巢蛋白,而有限数量的细胞(约占总细胞的 1%)表达神经元标记物 β-微管蛋白 III。这些结果总体上表明,EGF 固定的底物允许 iPS 细胞衍生的 NPC 有效增殖,同时保持未分化状态。基底上的大多数细胞(约占总细胞的 99%)表达 NPC 标记物巢蛋白,而有限数量的细胞(约占总细胞数的 1%)表达神经元标记物 β-微管蛋白 III。这些结果总体上表明,EGF 固定的底物允许 iPS 细胞衍生的 NPC 有效增殖,同时保持未分化状态。基底上的大多数细胞(约占总细胞的 99%)表达 NPC 标志物巢蛋白,而有限数量的细胞(约占总细胞的 1%)表达神经元标志物 β-微管蛋白 III。这些结果总体上表明,EGF 固定的底物允许 iPS 细胞衍生的 NPC 有效增殖,同时保持未分化状态。
更新日期:2020-05-30
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