Department of Cardiovascular Surgery, Osaka University, Graduate School of Medicine, Osaka, Japan.
Building Block Science Joint Research, Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
PLoS One. 2021 Mar 15;16(3):e0245571. doi: 10.1371/journal.pone.0245571. eCollection 2021.
The extracellular matrix (ECM) plays a key role in the viability and survival of implanted human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). We hypothesized that coating of three-dimensional (3D) cardiac tissue-derived hiPSC-CMs with the ECM protein fibronectin (FN) would improve the survival of transplanted cells in the heart and improve heart function in a rat model of ischemic heart failure. To test this hypothesis, we first explored the tolerance of FN-coated hiPSC-CMs to hypoxia in an in vitro study. For in vivo assessments, we constructed 3D-hiPSC cardiac tissues (3D-hiPSC-CTs) using a layer-by-layer technique, and then the cells were implanted in the hearts of a myocardial infarction rat model (3D-hiPSC-CTs, n = 10; sham surgery control group (without implant), n = 10). Heart function and histology were analyzed 4 weeks after transplantation. In the in vitro assessment, cell viability and lactate dehydrogenase assays showed that FN-coated hiPSC-CMs had improved tolerance to hypoxia compared with the control cells. In vivo, the left ventricular ejection fraction of hearts implanted with 3D-hiPSC-CT was significantly better than that of the sham control hearts. Histological analysis showed clear expression of collagen type IV and plasma membrane markers such as desmin and dystrophin in vivo after implantation of 3D-hiPSC-CT, which were not detected in 3D-hiPSC-CMs in vitro. Overall, these results indicated that FN-coated 3D-hiPSC-CT could improve distressed heart function in a rat myocardial infarction model with a well-expressed cytoskeletal or basement membrane matrix. Therefore, FN-coated 3D-hiPSC-CT may serve as a promising replacement for heart transplantation and left ventricular assist devices and has the potential to improve survivability and therapeutic efficacy in cases of ischemic heart disease.
细胞外基质 (ECM) 在植入的人诱导多能干细胞衍生的心肌细胞 (hiPSC-CMs) 的活力和存活中起着关键作用。我们假设，用 ECM 蛋白纤维连接蛋白 (FN) 涂覆三维 (3D) 心脏组织衍生的 hiPSC-CMs，将提高移植细胞在心脏中的存活率，并改善缺血性心力衰竭大鼠模型的心脏功能。为了验证这一假设，我们首先在体外研究中探索了 FN 涂覆的 hiPSC-CMs 对缺氧的耐受性。为了进行体内评估，我们使用逐层技术构建了 3D-hiPSC 心脏组织 (3D-hiPSC-CT)，然后将细胞植入心肌梗死大鼠模型的心脏中 (3D-hiPSC-CT，n = 10；假手术对照组（无植入），n = 10)。移植后 4 周分析心脏功能和组织学。在体外评估中，细胞活力和乳酸脱氢酶测定表明，与对照细胞相比，FN 涂覆的 hiPSC-CMs 对缺氧的耐受性有所提高。在体内，植入 3D-hiPSC-CT 的心脏的左心室射血分数明显优于假手术对照组的心脏。组织学分析显示，植入 3D-hiPSC-CT 后，体内可清晰表达胶原蛋白 IV 型和质膜标志物，如结蛋白和 dystrophin，而在体外的 3D-hiPSC-CMs 中则未检测到。总体而言，这些结果表明，FN 涂覆的 3D-hiPSC-CT 可改善大鼠心肌梗死模型中受损的心脏功能，且具有良好表达的细胞骨架或基底膜基质。因此，FN 涂覆的 3D-hiPSC-CT 可能成为心脏移植和左心室辅助装置的有前途的替代品，并有可能改善缺血性心脏病病例的存活率和治疗效果。
