Ohta Ryo, Niwa Akira, Taniguchi Yukimasa, Suzuki Naoya M, Toga Junko, Yagi Emiko, Saiki Norikazu, Nishinaka-Arai Yoko, Okada Chihiro, Watanabe Akira, Nakahata Tatsutoshi, Sekiguchi Kiyotoshi, Saito Megumu K
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, Suita, Japan.
Sci Rep. 2016 Nov 2;6:35680. doi: 10.1038/srep35680.
Obtaining highly purified differentiated cells via directed differentiation from human pluripotent stem cells (hPSCs) is an essential step for their clinical application. Among the various conditions that should be optimized, the precise role and contribution of the extracellular matrix (ECM) during differentiation are relatively unclear. Here, using a short fragment of laminin 411 (LM411-E8), an ECM predominantly expressed in the vascular endothelial basement membrane, we demonstrate that the directed switching of defined ECMs robustly yields highly-purified (>95%) endothelial progenitor cells (PSC-EPCs) without cell sorting from hPSCs in an integrin-laminin axis-dependent manner. Single-cell RNA-seq analysis revealed that LM411-E8 resolved intercellular transcriptional heterogeneity and escorted the progenitor cells to the appropriate differentiation pathway. The PSC-EPCs gave rise to functional endothelial cells both in vivo and in vitro. We therefore propose that sequential switching of defined matrices is an important concept for guiding cells towards desired fate.
通过人多能干细胞（hPSCs）的定向分化获得高度纯化的分化细胞是其临床应用的关键步骤。在各种需要优化的条件中，细胞外基质（ECM）在分化过程中的精确作用和贡献相对不清楚。在这里，我们使用层粘连蛋白411（LM411-E8）的一个短片段，一种主要在血管内皮基底膜中表达的细胞外基质，证明了特定细胞外基质的定向切换以整合素-层粘连蛋白轴依赖性方式，从hPSCs中无需细胞分选即可稳健地产生高度纯化（>95%）的内皮祖细胞（PSC-EPCs）。单细胞RNA测序分析表明，LM411-E8解决了细胞间转录异质性，并引导祖细胞走向适当的分化途径。PSC-EPCs在体内和体外都能产生功能性内皮细胞。因此，我们提出特定基质的顺序切换是引导细胞走向理想命运的一个重要概念。
