Generation of cardiac spheres from primate pluripotent stem cells in a small molecule-based 3D system
Generation of cardiac spheres from primate pluripotent stem cells in a small molecule-based 3D system
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在基于小分子的 3D 系统中利用灵长类多能干细胞生成心脏球体
DOI：
10.1016/j.biomaterials.2015.06.024
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发表时间：
2015-10-01
期刊：
影响因子：
14
通讯作者：
Li, Tianqing
中科院分区：
文献类型：
--
作者：
Jiang, Bin;Xiang, Zhen;Li, Tianqing
多能干细胞(PSC)在心脏再生医学中的应用需要在特定的培养液中培养出具有成熟表型的丰富的心肌细胞(CMS)。然而，目前的方法通常是在产生不成熟CMS的2D环境中执行的。在这里，我们报道了一种简单的、无生长因子的3D培养系统，可以使用3D培养的人和猴子的PSC球快速高效地产生85.07+/-1.8%的自发收缩心肌球(ScCDS)。结合基于小分子的3D诱导，该方法产生高达95.7%的CMS，每个输入多能细胞的产量高达237 CMS，对人和猴的PSCs有效，并在三个月以上保持81.03+/-12.43%的CDS的自发收缩。这些CDS显示了CM的超微结构、钙瞬变、适当的药理反应和成熟特有的CM基因表达谱。此外，3D来源的CMS比平行2D培养的CMS表现出更多的成熟表型。该系统适用于大规模生产用于疾病研究、细胞治疗和药物筛选的CMS。(C)2015爱思唯尔有限公司。保留所有权利。
Pluripotent stem cell (PSC) usage in heart regenerative medicine requires producing enriched cardiomyocytes (CMs) with mature phenotypes in a defined medium. However, current methods are typically performed in 2D environments that produce immature CMs. Here we report a simple, growth factor-free 3D culture system to rapidly and efficiently generate 85.07 +/- 1.8% of spontaneously contractile cardiac spheres (scCDSs) using 3D-cultured human and monkey PSC-spheres. Along with small molecule-based 3D induction, this protocol produces CDSs of up to 95.7% CMs at a yield of up to 237 CMs for every input pluripotent cell, is effective for human and monkey PSCs, and maintains 81.03 +/- 12.43% of CDSs in spontaneous contractibility for over three months. These CDSs displayed CM ultrastructure, calcium transient, appropriate pharmacological responses and CM gene expression profiles specific for maturity. Furthermore, 3D-derived CMs displayed more mature phenotypes than those from a parallel 2D-culture. The system is compatible to large-scaly produce CMs for disease study, cell therapy and pharmaceutics screening. (C) 2015 Elsevier Ltd. All rights reserved.