The myocardial regenerative potential of three-dimensional engineered cardiac tissues composed of multiple human iPS cell-derived cardiovascular cell lineages

Abstract

Human induced pluripotent stem cells (hiPSCs) are a robust source for cardiac regenerative therapy due to their potential to support autologous and allogeneic transplant paradigms. The in vitro generation of three-dimensional myocardial tissue constructs using biomaterials as an implantable hiPSC-derived myocardium provides a path to realize sustainable myocardial regeneration. We generated engineered cardiac tissues (ECTs) from three cellular compositions of cardiomyocytes (CMs), endothelial cells (ECs), and vascular mural cells (MCs) differentiated from hiPSCs. We then determined the impact of cell composition on ECT structural and functional properties. In vitro force measurement showed that CM+EC+MC ECTs possessed preferential electromechanical properties versus ECTs without vascular cells indicating that incorporation of vascular cells augmented tissue maturation and function. The inclusion of MCs facilitated more mature CM sarcomeric structure, preferential alignment, and activated multiple tissue maturation pathways. The CM+EC+MC ECTs implanted onto infarcted, immune tolerant rat hearts engrafted, displayed both host and graft-derived vasculature, and ameliorated myocardial dysfunction. Thus, a composition of CMs and multiple vascular lineages derived from hiPSCs and incorporated into ECTs promotes functional maturation and demonstrates myocardial replacement and perfusion relevant for clinical translation.

Impact of Cell Composition and Geometry on Human Induced Pluripotent Stem Cells-Derived Engineered Cardiac Tissue

“…We induced multiple CV cell lineages from hiPSCs to generate LF-ECTs using the lineage distribution shown to generate an optimal linear hiPSC-derived ECTs8. We employed two distinct CV cell differentiation protocols to generate either predominantly cardiac troponin-T (cTnT) + CMs and vascular endothelial (VE)-cadherin (CD144) + endothelial cells (ECs) or to generate predominantly platelet-derived growth factor receptor beta (PDGFRβ; CD140b) + vascular mural cells (MCs) (Fig.…”

“…Even with a broad range of evidence-based therapies, the five-year survival rate of heart failure remains as low as approximately 50%1. Numerous preclinical studies and clinical trials have suggested that application of stem and/or progenitor cell populations to an injured heart may hold a potential to ameliorate left ventricular dysfunction caused by ischemic and dilated cardiomyopathy accompanying heart failure2345678. Among various cell types, human induced pluripotent stem cells (hiPSCs) are considered highly promising cell sources for cardiac regenerative cell therapy, because the fundamental etiology of heart failure is the result of massive loss or dysfunction of myocardial cells9, and various cardiovascular (CV) cell lineages can be scalably produced from iPSCs101112.…”

“…Previously, we reported the generation of three-dimensional (3D) linear engineered cardiac tissues (ECTs) from chick embryonic or rat fetal cardiomyocytes (CMs) and biomaterials as a robust in vitro model to elucidate the development of embryonic myocardium and a platform to realise cardiac regeneration via implantation therapy for injured myocardium1819. In order to advance this technology towards clinical application, we developed and validated a method to generate linear ECTs from human iPSCs-derived CV lineages (hiPSC-ECTs)8. There we found that coexistence of multiple vascular lineages with CMs within the 3D ECT composition promoted structural and electrophysiological tissue maturation.…”

Untiring steps toward the maturation of human stem cell-engineered heart tissue

“…Another perspective is the supplementation of other cell lineages consisting native heart tissue such as vascular cells (vascular endothelial cells or vascular mural cells), or stromal cell lineages (e.g., cardiac fibroblasts) besides cardiomyocytes. Cell sheet experiments or 3-D engineered cardiac tissue experiments have shown that the co-existence of vascular cells with cardiomyocytes within the structure promotes reinforced secretion of humoral factors working on tissue repair, cellular alignment or sarcomeric maturation (7,19 (20). These results indicate the importance of co-existence of various cardiac cell lineages among the 3-D construct, and that the addition of electrical stimulation potentially further enhances tissue maturation.…”

“…Cardiac cells differentiated from hiPSCs mostly used for recent studies in this research field are juvenile and immature, and have only acquired characteristics of terminally differentiated cell lineages for 1-2 months. These cells are equivalent to cells at the fetal stage at the first trimester of pregnancy (7)(8)(9)(10). It has been reported that the electrophysiological properties of the vast majority of hiPSC-derived cardiomyocytes lack the I K1 current leading to immature repolarization capacity (16) which encourages researchers to drive cellular maturation, and also maturation as a tissue.…”