Scalable Production of Human Cardiac Microspheroids and Establishment of an In Vitro Diabetic Cardiomyopathy Model

Abstract

Recently, human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have emerged as a promising tool in regenerative medicine and disease modeling. Engineered cardiac tissues (ECTs) have received significant attention for their ability to provide a more native-like 3-dimensional (3D) environment compared to traditional 2-dimensional (2D) monolayer hiPSC-CMs. However, scaling up production and employing ECTs for disease modeling present challenges. This dissertation investigates various aspects of cardiac tissue engineering using hiPSCs to address the clinically relevant scale-up production of hiPSC-CMs and the creation of reliable, mature-like cardiac tissues for disease modeling and regenerative therapies. Project 1 focuses on comparing scaffold-based microsphere and aggregate platforms (which is the state-of-the- art approach for scaling up hiPSC-CM production) for efficient cardiac tissue production, demonstrating superior functionality and scalability of microspheres. Project 2 explores the influence of initial microenvironment geometry on hiPSC cardiac differentiation outcomes and maturity of hiPSC-CMs, emphasizing scalability and biomaterial encapsulation benefits. Project 3 examines strategies to enhance the metabolic maturation of ECTs, demonstrating significant improvements in structural, functionality, and metabolic maturity with employing maturation media (enriched fatty acids containing low glucose). Project 4 investigates the ability of mature ECTs to mimic certain pathological phenotypes of diabetic cardiomyopathy (DCMP), showcasing the potential use of this in vitro disease model for drug discovery. Collectively, these projects contribute to the advancement of cardiac tissue engineering, providing insights into scalability, maturation, and disease modeling for regenerative medicine and drug studies.