Spheroids of cardiomyocytes derived from human-induced pluripotent stem cells improve recovery from myocardial injury in mice
Spheroids of cardiomyocytes derived from human-induced pluripotent stem cells improve recovery from myocardial injury in mice
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DOI：
10.1152/ajpheart.00688.2017
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发表时间：
2018-08-01
影响因子：
4.8
通讯作者：
Zhang, Jianyi
中科院分区：
文献类型：
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作者：
Mattapally, Saidulu;Zhu, Wuqiang;Zhang, Jianyi
当心肌细胞在三维簇中培养时，天然心脏组织的微环境可以更好地复制（即，球状体）比单层或作为单个细胞。因此，我们在心肌细胞（hiPSC-CM）中分化人类心脏谱系诱导的多能干细胞，并使它们形成球状体和球状体融合体，这些球状体和球状体融合体在体外进行表征，并在实验诱导的心肌梗死（MI）后在小鼠中进行评估。在球体形成的24小时内观察到同步收缩，光学映射实验证实了Ca 2+瞬变和传播动作电位的存在。在球状体融合中，形成后第1-2天的球状体内传导速度为7.0 +/- 3.8 cm/s，而球状体之间的传导速度从第1-2天的0.8 +/- 1.1 cm/s显著增加（P = 0.003）至第7天的3.3 +/- 1.4 cm/s。对于鼠MI模型，将五个球状体融合体（200，000个hiPSC-CM/球状体）包埋在纤维蛋白贴片中，并将贴片移植到梗死部位上。后来（4周），超声心动图测量左心室射血分数和缩短分数显着更大的动物，在没有补丁恢复，移植率为25.6%或30%时，组织学或通过生物发光成像评价，分别。从球体补丁释放的外泌体似乎增加了心脏功能。总之，我们的结果确立了使用hiPSC-CM球状体和球状体融合体用于心脏组织工程的可行性，并且当在鼠MI模型中评价含有hiPSC-CM球状体融合体的纤维蛋白贴片时，植入率远高于我们通过直接心肌内注射实现的植入率。
The microenvironment of native heart tissue may be better replicated when cardiomyocytes are cultured in three-dimensional clusters (i.e.. spheroids) than in monolayers or as individual cells. Thus, we differentiated human cardiac lineage-induced pluripotent stem cells in cardiomyocytes (hiPSC-CMs) and allowed them to form spheroids and spheroid fusions that were characterized in vitro and evaluated in mice after experimentally induced myocardial infarction (MI). Synchronized contractions were observed within 24 h of spheroid formation, and optical mapping experiments confirmed the presence of both Ca2+ transients and propagating action potentials. In spheroid fusions, the intraspheroid conduction velocity was 7.0 +/- 3.8 cm/s on days 1-2 after formation, whereas the conduction velocity between spheroids increased significantly (P = 0.003) from 0.8 +/- 1.1 cm/s on days 1-2 to 3.3 +/- 1.4 cm/s on day 7. For the murine MI model, five-spheroid fusions (200,000 hiPSC-CMs/spheroid) were embedded in a fibrin patch and the patch was transplanted over the site of infarction. Later (4 wk), echocardiographic measurements of left ventricular ejection fraction and fractional shortening were significantly greater in patchtreated animals than in animals that recovered without the patch, and the engraftment rate was 25.6% or 30% when evaluated histologically or via bioluminescence imaging, respectively. The exosomes released from the spheroid patch seemed to increase cardiac function. In conclusion, our results established the feasibility of using hiPSC-CM spheroids and spheroid fusions for cardiac tissue engineering, and, when fibrin patches containing hiPSC-CM spheroid fusions were evaluated in a murine MI model, the engraftment rate was much higher than the rates we have achieved via the direct intramyocardial injection.