Induced Pluripotent Stem Cell-Derived Cardiomyocytes Provide In Vivo Biological Pacemaker Function.
Induced Pluripotent Stem Cell-Derived Cardiomyocytes Provide In Vivo Biological Pacemaker Function.
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DOI：
10.1161/circep.116.004508
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发表时间：
2017-05
期刊：
影响因子：
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通讯作者：
Rosen MR
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作者：
Chauveau S;Anyukhovsky EP;Ben-Ari M;Naor S;Jiang YP;Danilo P Jr;Rahim T;Burke S;Qiu X;Potapova IA;Doronin SV;Brink PR;Binah O;Cohen IS;Rosen MR
尽管已经使用多种方法在动物模型中创建生物起搏器，但尚未为此目的研究诱导多能干细胞衍生的心肌细胞（iPSC-CM）。我们现在报告iPSC-CM在犬模型中的起搏器功能。拟胚体来源于人角质形成细胞，确定其动作电位特征，并鉴定其基因表达谱和分化标志物。房室传导阻滞犬，免疫抑制后，植入VVI起搏器，心外膜下注射40 ~ 75个节律性收缩的拟胚体（共1.3 ~ 2×106个细胞）于左室前基底部。每两周进行一次ECG和24小时霍尔特监测。4 ~ 13周后，输注肾上腺素（1 μg kg−1 min−1），并取出心脏进行组织学或电生理学研究。iPSC-CM在很大程度上失去了多能性标志物，对心脏特异性标志物呈阳性。并表现出If依赖性自动性。植入后第1周，注射部位的心外膜起搏识别出该部位产生的匹配搏动。到第4周，20%的心跳是电子起搏的，60%到80%的心跳是匹配的，平均和最大生物起搏器频率分别为45和75次/分钟。第4周时，匹配搏动的夜间和日间最大频率分别为53±6.9和69±10.4次/分钟。肾上腺素可使匹配心跳频率从35±4.3次/min增加到65±4.0次/min。用活体染料Dil孵育胚状体，显示注射细胞在给药部位的持久性。iPSC-CM可以整合到宿主心肌中并产生生物起搏器。虽然这是一个有希望的发展，但iPSC-CM起搏器的频率和节奏仍有待优化。
Although multiple approaches have been used to create biological pacemakers in animal models, induced pluripotent stem cell–derived cardiomyocytes (iPSC-CMs) have not been investigated for this purpose. We now report pacemaker function of iPSC-CMs in a canine model. Embryoid bodies were derived from human keratinocytes, their action potential characteristics determined, and their gene expression profiles and markers of differentiation identified. Atrioventricular blocked dogs were immunosuppressed, instrumented with VVI pacemakers, and injected subepicardially into the anterobasal left ventricle with 40 to 75 rhythmically contracting embryoid bodies (totaling 1.3–2×106 cells). ECG and 24-hour Holter monitoring were performed biweekly. After 4 to 13 weeks, epinephrine (1 μg kg−1 min−1) was infused, and the heart removed for histological or electrophysiological study. iPSC-CMs largely lost the markers of pluripotency, became positive for cardiac-specific markers. and manifested If-dependent automaticity. Epicardial pacing of the injection site identified matching beats arising from that site by week 1 after implantation. By week 4, 20% of beats were electronically paced, 60% to 80% of beats were matching, and mean and maximal biological pacemaker rates were 45 and 75 beats per minute. Maximum night and day rates of matching beats were 53±6.9 and 69±10.4 beats per minute, respectively, at 4 weeks. Epinephrine increased rate of matching beats from 35±4.3 to 65±4.0 beats per minute. Incubation of embryoid bodies with the vital dye, Dil, revealed the persistence of injected cells at the site of administration. iPSC-CMs can integrate into host myocardium and create a biological pacemaker. Although this is a promising development, rate and rhythm of the iPSC-CMs pacemakers remain to be optimized.