Bai Xiaowen, Pinkernell Kai, Song Yao-Hua, Nabzdyk Christoph, Reiser Jakob, Alt Eckhard
Department of Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA.
Biochem Biophys Res Commun. 2007 Feb 16;353(3):665-71. doi: 10.1016/j.bbrc.2006.12.103. Epub 2006 Dec 22.
In the present study, the potential of human adipose-derived stem cells to differentiate into cells with characteristics of cardiomyocytes was investigated. Adipose tissue-derived stem cells (ADSCs) were transduced with two different lentiviral vectors simultaneously: (1) a lentiviral vector expressing eGFP controlled by the Nkx2.5 promoter and (2) a lentiviral vector expressing DsRed2 controlled by the myosin light chain-2v promoter (MLC-2v). Nkx2.5-eGFP and MLC-2v-DsRed2 dual positive cells were isolated by FACS. Immunostaining and RT-PCR analysis of the dual positive cells revealed that these cells are positive for Nkx2.5, cardiac troponin I, and L-type calcium channel alpha-1c subunit. Electrophysiology studies demonstrated the presence of functional voltage-dependent calcium and potassium channels. These observations confirm that cardiac progenitor cells can be isolated and enriched from human adipose-derived stem cells using lentiviral selection, and they might represent a new source for cell therapy for myocardial infarction and heart failure.
在本研究中,对人脂肪来源干细胞分化为具有心肌细胞特征的细胞的潜力进行了研究。脂肪组织来源干细胞(ADSCs)同时用两种不同的慢病毒载体进行转导:(1)一种由Nkx2.5启动子控制表达增强绿色荧光蛋白(eGFP)的慢病毒载体,以及(2)一种由肌球蛋白轻链-2v启动子(MLC-2v)控制表达红色荧光蛋白(DsRed2)的慢病毒载体。通过荧光激活细胞分选(FACS)分离出Nkx2.5-eGFP和MLC-2v-DsRed2双阳性细胞。对双阳性细胞进行免疫染色和逆转录-聚合酶链反应(RT-PCR)分析显示,这些细胞对Nkx2.5、心肌肌钙蛋白I和L型钙通道α-1c亚基呈阳性。电生理学研究证明存在功能性电压依赖性钙通道和钾通道。这些观察结果证实,利用慢病毒筛选可从人脂肪来源干细胞中分离和富集心脏祖细胞,它们可能代表心肌梗死和心力衰竭细胞治疗的新来源。