Tashiro Katsuhisa, Hirata Nobue, Okada Atsumasa, Yamaguchi Tomoko, Takayama Kazuo, Mizuguchi Hiroyuki, Kawabata Kenji
Laboratory of Stem Cell Regulation and Laboratory of Hepatocyte Regulation, National Institute of Biomedical Innovation, Osaka, Japan; Laboratory of Biochemistry and Molecular Biology, iPS Cell-Based Research Project on Hepatic Toxicity and Metabolism, and Laboratory of Biomedical Innovation, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; The Center for Advanced Medical Engineering and Informatics, Osaka University, Osaka, Japan.
Laboratory of Stem Cell Regulation and Laboratory of Hepatocyte Regulation, National Institute of Biomedical Innovation, Osaka, Japan; Laboratory of Biochemistry and Molecular Biology, iPS Cell-Based Research Project on Hepatic Toxicity and Metabolism, and Laboratory of Biomedical Innovation, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; The Center for Advanced Medical Engineering and Informatics, Osaka University, Osaka, Japan
Stem Cells Transl Med. 2015 May;4(5):424-36. doi: 10.5966/sctm.2014-0173. Epub 2015 Mar 11.
In developing embryos or in vitro differentiation cultures using pluripotent stem cells (PSCs), such as embryonic stem cells and induced pluripotent stem cells, fetal liver kinase 1 (Flk1)-expressing mesodermal cells are thought to be a heterogeneous population that includes hematopoietic progenitors, endothelial progenitors, and cardiac progenitors. However, information on cell surface markers for separating these progenitors in Flk1⁺ cells is currently limited. In the present study, we show that distinct types of progenitor cells in Flk1⁺ cells could be separated according to the expression of coxsackievirus and adenovirus receptor (CAR, also known as CXADR), a tight junction component molecule. We found that mouse and human PSC- and mouse embryo-derived Flk1⁺ cells could be subdivided into Flk1⁺CAR⁺ cells and Flk1⁺CAR⁻ cells. The progenitor cells with cardiac potential were almost entirely restricted to Flk1⁺CAR⁺ cells, and Flk1⁺CAR⁻ cells efficiently differentiated into hematopoietic cells. Endothelial differentiation potential was observed in both populations. Furthermore, from the expression of CAR, Flk1, and platelet-derived growth factor receptor-α (PDGFRα), Flk1⁺ cells could be separated into three populations (Flk1⁺PDGFRα⁻ CAR⁻ cells, Flk1⁺PDGFRα⁻CAR⁺ cells, and Flk1⁺PDGFRα⁺CAR⁺ cells). Flk1⁺PDGFRα⁺ cells and Flk1⁺PDGFRα⁻ cells have been reported as cardiac and hematopoietic progenitor cells, respectively. We identified a novel population (Flk1⁺PDGFRα⁻ CAR⁺ cells) with the potential to differentiate into not only hematopoietic cells and endothelial cells but also cardiomyocytes. Our findings indicate that CAR would be a novel and prominent marker for separating PSC- and embryo-derived Flk1⁺ mesodermal cells with distinct differentiation potentials.
在发育中的胚胎或使用多能干细胞（PSC）（如胚胎干细胞和诱导多能干细胞）进行的体外分化培养中，表达胎儿肝激酶1（Flk1）的中胚层细胞被认为是一个异质群体，包括造血祖细胞、内皮祖细胞和心脏祖细胞。然而，目前关于在Flk1⁺细胞中分离这些祖细胞的细胞表面标志物的信息有限。在本研究中，我们表明，Flk1⁺细胞中不同类型的祖细胞可以根据柯萨奇病毒和腺病毒受体（CAR，也称为CXADR）的表达进行分离，CAR是一种紧密连接成分分子。我们发现，小鼠和人类PSC以及小鼠胚胎来源的Flk1⁺细胞可细分为Flk1⁺CAR⁺细胞和Flk1⁺CAR⁻细胞。具有心脏分化潜能的祖细胞几乎完全局限于Flk1⁺CAR⁺细胞，而Flk1⁺CAR⁻细胞则有效地分化为造血细胞。在这两个群体中均观察到内皮分化潜能。此外，根据CAR、Flk1和血小板衍生生长因子受体-α（PDGFRα）的表达，Flk1⁺细胞可分为三个群体（Flk1⁺PDGFRα⁻CAR⁻细胞、Flk1⁺PDGFRα⁻CAR⁺细胞和Flk1⁺PDGFRα⁺CAR⁺细胞）。Flk1⁺PDGFRα⁺细胞和Flk1⁺PDGFRα⁻细胞分别被报道为心脏和造血祖细胞。我们鉴定出一个新的群体（Flk1⁺PDGFRα⁻CAR⁺细胞），其不仅具有分化为造血细胞和内皮细胞的潜能，还具有分化为心肌细胞的潜能。我们的研究结果表明，CAR将是一种用于分离具有不同分化潜能的PSC和胚胎来源的Flk1⁺中胚层细胞的新型且重要的标志物。
