Department of Dermatology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, CO, USA.
Charles C. Gates Center for Regenerative Medicine, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, CO, USA.
Methods Mol Biol. 2022;2549:169-186. doi: 10.1007/7651_2021_377.
Endothelial cells (ECs) are important components of the circulatory system. These cells can be used for in vitro modeling of cardiovascular diseases and in regenerative medicine to promote vascularization of engineered tissue constructs. However, low proliferative capacity and patient-to-patient variability limit the use of primary ECs in the clinic and disease modeling. ECs differentiated from human induced pluripotent stem cells (iPSCs) can serve as a viable alternative to primary ECs for these applications. This is because human iPSCs can proliferate indefinitely and have the potential to differentiate into a variety of somatic cell lines, providing a renewable source of patient-specific cells. Here, we present an optimized, highly reproducible method for the differentiation of human iPSCs toward vascular ECs. The protocol relies on the activation of the WNT signaling pathway and the use of growth factors and small molecules. The resulting iPSC-derived ECs can be cultured for multiple passages without losing their functionality and are suitable for both in vitro and in vivo studies.
内皮细胞(ECs)是循环系统的重要组成部分。这些细胞可用于体外心血管疾病建模和再生医学,以促进工程组织构建体的血管生成。然而,低增殖能力和患者间的可变性限制了原代 ECs 在临床和疾病建模中的应用。从人诱导多能干细胞(iPSCs)分化而来的 ECs 可作为这些应用中原代 ECs 的可行替代品。这是因为人 iPSCs 可以无限增殖,并且有可能分化为多种体细胞系,提供了可再生的患者特异性细胞来源。在这里,我们提出了一种优化的、高度可重复的方法,用于将人 iPSCs 分化为血管内皮细胞。该方案依赖于 WNT 信号通路的激活以及生长因子和小分子的使用。由此产生的 iPSC 衍生的 ECs 可以在不丧失其功能的情况下进行多次传代培养,适用于体外和体内研究。