Jumabay Medet, Matsumoto Taro, Yokoyama Shin-ichiro, Kano Koichiro, Kusumi Yoshiaki, Masuko Takayuki, Mitsumata Masako, Saito Satoshi, Hirayama Atsushi, Mugishima Hideo, Fukuda Noboru
Division of Cardiology, Department of Medicine, Nihon University School of Medicine, Tokyo 173-8610, Japan.
J Mol Cell Cardiol. 2009 Nov;47(5):565-75. doi: 10.1016/j.yjmcc.2009.08.004. Epub 2009 Aug 15.
Adipose tissue-derived stem cells have been demonstrated to differentiate into cardiomyocytes and vascular endothelial cells. Here we investigate whether mature adipocyte-derived dedifferentiated fat (DFAT) cells can differentiate to cardiomyocytes in vitro and in vivo by establishing DFAT cell lines via ceiling culture of mature adipocytes. DFAT cells were obtained by dedifferentiation of mature adipocytes from GFP-transgenic rats. We evaluated the differentiating ability of DFAT cells into cardiomyocytes by detection of the cardiac phenotype markers in immunocytochemical and RT-PCR analyses in vitro. We also examined effects of the transplantation of DFAT cells into the infarcted heart of rats on cardiomyocytes regeneration and angiogenesis. DFAT cells expressed cardiac phenotype markers when cocultured with cardiomyocytes and also when grown in MethoCult medium in the absence of cardiomyocytes, indicating that DFAT cells have the potential to differentiate to cardiomyocyte lineage. In a rat acute myocardial infarction model, transplanted DFAT cells were efficiently accumulated in infarcted myocardium and expressed cardiac sarcomeric actin at 8 weeks after the cell transplantation. The transplantation of DFAT cells significantly (p<0.05) increased capillary density in the infarcted area when compared with hearts from saline-injected control rats. We demonstrated that DFAT cells have the ability to differentiate to cardiomyocyte-like cells in vitro and in vivo. In addition, transplantation of DFAT cells led to neovascuralization in rats with myocardial infarction. We propose that DFAT cells represent a promising candidate cell source for cardiomyocyte regeneration in severe ischemic heart disease.
脂肪组织来源的干细胞已被证明可分化为心肌细胞和血管内皮细胞。在此，我们通过成熟脂肪细胞的悬滴培养建立去分化脂肪（DFAT）细胞系，来研究成熟脂肪细胞来源的DFAT细胞在体外和体内是否能分化为心肌细胞。DFAT细胞是通过对绿色荧光蛋白转基因大鼠的成熟脂肪细胞进行去分化而获得的。我们通过体外免疫细胞化学和逆转录-聚合酶链反应（RT-PCR）分析中检测心脏表型标志物，来评估DFAT细胞向心肌细胞的分化能力。我们还研究了将DFAT细胞移植到大鼠梗死心脏中对心肌细胞再生和血管生成的影响。DFAT细胞与心肌细胞共培养时，以及在没有心肌细胞的情况下在甲基纤维素培养基中生长时，均表达心脏表型标志物，这表明DFAT细胞具有分化为心肌细胞谱系的潜力。在大鼠急性心肌梗死模型中，移植的DFAT细胞在细胞移植后8周有效地积聚在梗死心肌中，并表达心脏肌节肌动蛋白。与注射生理盐水的对照大鼠的心脏相比，DFAT细胞移植显著（p<0.05）增加了梗死区域的毛细血管密度。我们证明DFAT细胞在体外和体内都有能力分化为心肌样细胞。此外，DFAT细胞移植导致心肌梗死大鼠出现新生血管形成。我们认为，DFAT细胞是严重缺血性心脏病中心肌细胞再生的一个有前景的候选细胞来源。
