Kosair Charities Pediatric Heart Research Program, Cardiovascular Innovation Institute, University of Louisville, Louisville, Kentucky, The United States of America.
Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Sci Rep. 2017 Apr 3;7:45641. doi: 10.1038/srep45641.
The current study describes a scalable, porous large-format engineered cardiac tissue (LF-ECT) composed of human induced pluripotent stem cells (hiPSCs) derived multiple lineage cardiac cells with varied 3D geometries and cell densities developed towards the goal of scale-up for large animal pre-clinical studies. We explored multiple 15 × 15 mm ECT geometries using molds with rectangular internal staggered posts (mesh, ME), without posts (plain sheet, PS), or long parallel posts (multiple linear bundles, ML) and a gel matrix containing hiPSC-derived cardiomyocytes, endothelial, and vascular mural cells matured in vitro for 14 days. ME-ECTs displayed the lowest dead cell ratio (p < 0.001) and matured into 0.5 mm diameter myofiber bundles with greater 3D cell alignment and higher active stress than PS-ECTs. Increased initial ECT cell number beyond 6 M per construct resulted in reduced cell survival and lower active stress. The 6M-ME-ECTs implanted onto 1 week post-infarct immune tolerant rat hearts engrafted, displayed evidence for host vascular coupling, and recovered myocardial structure and function with reduced scar area. We generated a larger (30 × 30 mm) ME-ECT to confirm scalability. Thus, large-format ECTs generated from hiPSC-derived cardiac cells may be feasible for large animal preclinical cardiac regeneration paradigms.
当前的研究描述了一种可扩展的多孔大型工程化心脏组织 (LF-ECT),由源自人诱导多能干细胞 (hiPSC) 的多谱系心脏细胞组成,具有不同的 3D 几何形状和细胞密度,旨在为大型动物临床前研究的规模化发展。我们使用具有矩形内部交错柱的模具探索了多种 15×15mm 的 ECT 几何形状(带有网格的 ME、无柱的 PS 或长平行柱的 ML)和含有 hiPSC 衍生的心肌细胞、内皮细胞和血管周细胞的凝胶基质,这些细胞在体外成熟 14 天。ME-ECT 显示出最低的死细胞比例(p<0.001),并成熟为 0.5mm 直径的肌纤维束,具有更好的 3D 细胞排列和比 PS-ECT 更高的主动应力。每个构建体中超过 6M 的初始 ECT 细胞数量增加会导致细胞存活率降低和主动应力降低。将 6M-ME-ECT 植入 1 周后梗死的免疫耐受大鼠心脏中,显示出与宿主血管耦联的证据,并恢复了心肌结构和功能,减少了疤痕面积。我们生成了一个更大的 (30×30mm) ME-ECT 以确认可扩展性。因此,源自 hiPSC 衍生的心脏细胞的大型 ECT 可能适用于大型动物临床前心脏再生范式。