Jahan Basharat, McCloskey Kara E
Graduate Program in Biological Engineering and Small-scale Technologies, University of California, Merced, United States.
Graduate Program in Biological Engineering and Small-scale Technologies, University of California, Merced, United States; Department of Materials Science and Engineering, University of California, Merced, United States.
Stem Cell Res. 2020 Jan;42:101685. doi: 10.1016/j.scr.2019.101685. Epub 2019 Dec 16.
Human endothelial cells (ECs) are important tools in research and development of new therapies in the fields of angiogenesis, vasculogenesis, engineering organoids and multicellular tissues, drug discovery, and disease modeling. Efficient and robust induction of ECs from human pluripotent stem cells (hPSCs) serve as a renewable and indefinite cell sources. However, individual lines of embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are distinct and can often respond very differently to the same microenvironmental cues. Therefore, we set out to develop a differentiation methodology specifically designed for robustness across multiple human iPSC lines. In general, the key soluble signals remain consistent across cell lines, but because the differentiation and proliferation kinetics can differ slightly in hESC and iPSC cell lines, the time point for KDR+ cell sorting must be pre-determined for each cell line. This three-stage induction method uses three different chemically defined medium formulations and generates highly purified populations of actively proliferating and functional VE-cadherin+ ECs within 30 days.
人内皮细胞(ECs)是血管生成、血管发生、工程类器官和多细胞组织、药物发现及疾病建模等领域新疗法研发中的重要工具。从人多能干细胞(hPSCs)高效且稳健地诱导生成ECs可作为一种可再生且无限的细胞来源。然而,各胚胎干细胞系(hESCs)和诱导多能干细胞系(iPSCs)是不同的,并且对相同的微环境信号常常会有非常不同的反应。因此,我们着手开发一种专门为跨多个人iPSC系实现稳健性而设计的分化方法。一般来说,关键的可溶性信号在各细胞系中保持一致,但由于hESC和iPSC细胞系中的分化和增殖动力学可能略有不同,必须为每个细胞系预先确定KDR+细胞分选的时间点。这种三阶段诱导方法使用三种不同的化学成分确定的培养基配方,并在30天内生成高度纯化的、处于活跃增殖状态且具有功能的VE-钙黏蛋白+ ECs群体。