Microfluidic Single-Cell Analysis of Transplanted Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes After Acute Myocardial Infarction.
Microfluidic Single-Cell Analysis of Transplanted Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes After Acute Myocardial Infarction.
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DOI：
10.1161/circulationaha.114.015231
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发表时间：
2015-08-25
期刊：
影响因子：
37.8
通讯作者：
Wu JC
中科院分区：
文献类型：
--
作者：
Ong SG;Huber BC;Lee WH;Kodo K;Ebert AD;Ma Y;Nguyen PK;Diecke S;Chen WY;Wu JC
人类诱导多能干细胞（iPSC）是治疗用途的有吸引力的候选者，具有替代缺陷细胞和改善损伤或疾病环境中的功能恢复的潜力。在这里，我们测试的假设，即人iPSC衍生的心肌细胞（iPSC-CMs）可以分泌细胞因子作为分子基础，以减轻心肌梗死（MI）后的不良心脏重塑。人iPSC由皮肤成纤维细胞产生，并使用基于小分子的方案在体外分化。肌钙蛋白+iPSC-CM通过免疫组织化学、定量PCR、荧光激活细胞分选（FACS）和电生理学测量确认。随后，将来自表达萤火虫荧光素酶和GFP的载体转导的细胞系的2×106个iPSC-CM移植到急性左前降支（LAD）结扎的成年NOD/SCID小鼠体内。对照动物接受PBS注射。生物发光成像（BLI）显示移植到缺血心肌后植入有限。然而，与PBS处理的对照动物相比，移植iPSC-CM的动物的磁共振成像（MRI）在第35天显示出显著的功能改善和减弱的心脏重塑（射血分数：24.5±1.3对14.5±1.5%; P<0.05）。为了理解潜在的分子机制，使用收获的iPSC-CM的微流体单细胞分析、宿主心肌的激光捕获显微切割（LCM）和体外缺血刺激来证明iPSC-CM可以在缺血微环境中释放显著水平的促血管生成因子和抗凋亡因子。将人iPSC-CM移植到急性小鼠MI模型中可以改善左心室功能并减弱心脏重构。由于有限的植入，大多数的影响可能是解释这些细胞的旁分泌活动。
Human induced pluripotent stem cells (iPSCs) are attractive candidates for therapeutic use, with the potential to replace deficient cells and to improve functional recovery in injury or disease settings. Here we test the hypothesis that human iPSC-derived cardiomyocytes (iPSC-CMs) can secrete cytokines as a molecular basis to attenuate adverse cardiac remodeling after myocardial infarction (MI). Human iPSCs were generated from skin fibroblasts and differentiated in vitro using a small molecule based protocol. Troponin+ iPSC-CMs were confirmed by immunohistochemistry, quantitative PCR, fluorescence activated cell sorting (FACS), and electrophysiological measurements. Afterwards, 2×106 iPSC-CMs derived from a cell line transduced with a vector expressing firefly luciferase and GFP were transplanted into adult NOD/SCID mice with acute left anterior descending (LAD) ligation. Control animals received PBS injection. Bioluminescence imaging (BLI) showed limited engraftment upon transplantation into ischemic myocardium. However, magnetic resonance imaging (MRI) of animals transplanted with iPSC-CMs showed significant functional improvement and attenuated cardiac remodeling when compared to PBS-treated control animals at day 35 (Ejection fraction: 24.5±1.3 vs. 14.5±1.5%; P<0.05). To understand the underlying molecular mechanism, microfluidic single cell profiling of harvested iPSC-CMs, laser capture microdissection (LCM) of host myocardium, and in vitro ischemia stimulation were used to demonstrate that the iPSC-CMs could release significant levels of pro-angiogenic and anti-apoptotic factors in the ischemic microenvironment. Transplantation of human iPSC-CMs into an acute mouse MI model can improve left ventricular function and attenuate cardiac remodeling. Because of limited engraftment, most of the effects are possibly explained by paracrine activity of these cells.