Hirata Hirokazu, Kawamata Shin, Murakami Yoshinobu, Inoue Kayoko, Nagahashi Ayako, Tosaka Mako, Yoshimura Naoko, Miyamoto Yoshiaki, Iwasaki Hiroto, Asahara Takayuki, Sawa Yoshiki
Department of Tissue Engineering and Cell Therapy, Foundation of Biomedical Research and Innovation, 2-2 Minatojima-Minamimachi, Kobe, Japan.
J Biosci Bioeng. 2007 May;103(5):412-9. doi: 10.1263/jbb.103.412.
Nascent mesodermal cells derived from EB5 embryonic stem (ES) cells were sorted in terms of cardiogenic potential on the basis of their expression levels of platelet-derived growth factor receptor alpha (PDGFRalpha) and fetal liver kinase 1 (Flk-1). The sorted cells were cocultured with OP9 stromal cells to induce terminal differentiation into contractile cardiac colonies. A significant number of cardiac colonies were found in the Flk-1+/PDGFRalpha+ fraction. The enrichment double-positive fraction produced approximately fivefold more cardiac colonies than the Flk-1+/PDGFRalpha- fraction and 10-fold more than the Flk-1-/PDGFRalpha+ fraction. To investigate the involvement of these markers in embryonic cardiogenesis, the cells that disseminated from the E7.5-7.75 embryos were fractionated and seeded on OP9 cells. The cardiogenic potential was markedly enhanced in the Flk-1+/PDGFRalpha+ fraction. These results suggest that some of the precursor cells coexpressing these markers are selectively involved in cardiogenic events, and that the identification of ES-cell-derived precursors with these markers will contribute to the effective production of cardiomyocytes for cell therapies.
从EB5胚胎干细胞衍生的新生中胚层细胞，根据其血小板衍生生长因子受体α（PDGFRα）和胎儿肝激酶1（Flk-1）的表达水平，按照心脏发生潜能进行分选。将分选后的细胞与OP9基质细胞共培养，以诱导其终末分化为收缩性心脏集落。在Flk-1+/PDGFRα+组分中发现了大量心脏集落。富集的双阳性组分产生的心脏集落比Flk-1+/PDGFRα-组分多约五倍，比Flk-1-/PDGFRα+组分多十倍。为了研究这些标志物在胚胎心脏发生中的作用，将从E7.5 - 7.75胚胎中分散出来的细胞进行分级分离，并接种到OP9细胞上。Flk-1+/PDGFRα+组分的心脏发生潜能显著增强。这些结果表明，一些共表达这些标志物的前体细胞选择性地参与心脏发生事件，并且用这些标志物鉴定胚胎干细胞衍生的前体细胞将有助于为细胞治疗有效生产心肌细胞。
