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无动物培养体系和二维生物材料上 hPSC 的增殖。

Xeno-free culture and proliferation of hPSCs on 2D biomaterials.

机构信息

State Key Laboratory of Ophthalmology, Optometry and Visual Science, Eye Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China.

Department of Chemical and Materials Engineering, National Central University, Jhongli, Taoyuan, Taiwan.

出版信息

Prog Mol Biol Transl Sci. 2023;199:63-107. doi: 10.1016/bs.pmbts.2023.02.008. Epub 2023 Mar 14.

Abstract

Human pluripotent stem cells (human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs)) have unlimited proliferative potential, whereas adult stem cells such as bone marrow-derived stem cells and adipose-derived stem cells have problems with aging. When hPSCs are intended to be cultured on feeder-free or xeno-free conditions without utilizing mouse embryonic fibroblasts or human fibroblasts, they cannot be cultured on conventional tissue culture polystyrene dishes, as adult stem cells can be cultured but should be cultivated on material surfaces grafted or coated with (a) natural or recombinant extracellular matrix (ECM) proteins, (b) ECM protein-derived peptides and specific synthetic polymer surfaces in xeno-free and/or chemically defined conditions. This review describes current developing cell culture biomaterials for the proliferation of hPSCs while maintaining the pluripotency and differentiation potential of the cells into 3 germ layers. Biomaterials for the cultivation of hPSCs without utilizing a feeder layer are essential to decrease the risk of xenogenic molecules, which contributes to the potential clinical usage of hPSCs. ECM proteins such as human recombinant vitronectin, laminin-511 and laminin-521 have been utilized instead of Matrigel for the feeder-free cultivation of hPSCs. The following biomaterials are also discussed for hPSC cultivation: (a) decellularized ECM, (b) peptide-grafted biomaterials derived from ECM proteins, (c) recombinant E-cadherin-coated surface, (d) polysaccharide-immobilized surface, (e) synthetic polymer surfaces with and without bioactive sites, (f) thermoresponsive polymer surfaces with and without bioactive sites, and (g) synthetic microfibrous scaffolds.

摘要

人多能干细胞(人胚胎干细胞(hESCs)和诱导多能干细胞(hiPSCs))具有无限的增殖潜力,而骨髓源性干细胞和脂肪源性干细胞等成体干细胞则存在老化问题。当 hPSCs 旨在在无饲养层或无异种条件下培养,而不利用小鼠胚胎成纤维细胞或人成纤维细胞时,它们不能在常规的组织培养聚苯乙烯培养皿上培养,而成体干细胞可以培养,但应在材料表面上进行培养,该表面接枝或涂覆有(a)天然或重组细胞外基质(ECM)蛋白,(b)ECM 蛋白衍生肽和特定的合成聚合物表面,这些表面在无异种和/或化学定义条件下。本综述描述了当前用于 hPSC 增殖的细胞培养生物材料的发展情况,同时保持了细胞向 3 个胚层分化的多能性和分化潜能。在不利用饲养层的情况下培养 hPSC 的生物材料对于降低异种分子的风险至关重要,这有助于 hPSC 的潜在临床应用。已经利用人重组 vitronectin、层粘连蛋白-511 和层粘连蛋白-521 等 ECM 蛋白替代 Matrigel 用于 hPSC 的无饲养层培养。还讨论了用于 hPSC 培养的以下生物材料:(a)脱细胞 ECM,(b)源自 ECM 蛋白的肽接枝生物材料,(c)重组 E-钙粘蛋白涂覆表面,(d)多糖固定表面,(e)具有和不具有生物活性位点的合成聚合物表面,(f)具有和不具有生物活性位点的热响应聚合物表面,以及(g)合成微纤维支架。

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