Kado Miwako, Lee Jong-Kook, Hidaka Kyoko, Miwa Keiko, Murohara Toyoaki, Kasai Kenji, Saga Shinsuke, Morisaki Takayuki, Ueda Yuichi, Kodama Itsuo
Department of Cardiothoracic Surgery, Nagoya University, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan.
Biochem Biophys Res Commun. 2008 Dec 12;377(2):413-418. doi: 10.1016/j.bbrc.2008.09.160. Epub 2008 Oct 18.
For myocardial regeneration therapy, the low differentiation capability of functional cardiomyocytes sufficient to replace the damaged myocardial tissue is one of the major difficulties. Using Nkx2.5-GFP knock-in ES cells, we show a new efficient method to obtain cardiomyocytes from embryonic stem (ES) cells. The proportion of GFP-positive cells was significantly increased when ES cells were cultured with a conditioned medium from aortic endothelial cells (ECs), accompanied by upregulation of cardiac-specific genes as well as other mesodermal genes. The promotion was more prominent when EC-conditioned medium was added at an early stage of ES cell differentiation culture (Day 0-3). Inhibitors of bone morphogenic protein (BMP), cyclooxygenase (COX), and nitric oxide synthetase (NO) prevented the promotion of cardiomyogenesis by EC-conditioned medium. These results suggest that supplementation of EC-conditioned medium enables cardiomyocytes to be obtained efficiently through promotion of mesoderm induction, which is regulated by BMP, COX, and NOS.
对于心肌再生治疗而言，功能性心肌细胞分化能力低下，不足以替代受损心肌组织，是主要困难之一。利用Nkx2.5-GFP基因敲入胚胎干细胞（ES细胞），我们展示了一种从胚胎干细胞中获取心肌细胞的新的有效方法。当ES细胞与主动脉内皮细胞（ECs）的条件培养基一起培养时，绿色荧光蛋白（GFP）阳性细胞的比例显著增加，同时心脏特异性基因以及其他中胚层基因上调。当在ES细胞分化培养的早期阶段（第0 - 3天）添加EC条件培养基时，这种促进作用更为显著。骨形态发生蛋白（BMP）、环氧化酶（COX）和一氧化氮合酶（NO）的抑制剂可阻止EC条件培养基对心肌生成的促进作用。这些结果表明，补充EC条件培养基能够通过促进由BMP, COX和NOS调节的中胚层诱导，有效地获得心肌细胞。
