From Human Pluripotent Stem Cells to 3D Cardiac Microtissues: Progress, Applications and Challenges

Abstract

The knowledge acquired throughout the years concerning the in vivo regulation of cardiac development has promoted the establishment of directed differentiation protocols to obtain cardiomyocytes (CMs) and other cardiac cells from human pluripotent stem cells (hPSCs), which play a crucial role in the function and homeostasis of the heart. Among other developments in the field, the transition from homogeneous cultures of CMs to more complex multicellular cardiac microtissues (MTs) has increased the potential of these models for studying cardiac disorders in vitro and for clinically relevant applications such as drug screening and cardiotoxicity tests. This review addresses the state of the art of the generation of different cardiac cells from hPSCs and the impact of transitioning CM differentiation from 2D culture to a 3D environment. Additionally, current methods that may be employed to generate 3D cardiac MTs are reviewed and, finally, the adoption of these models for in vitro applications and their adaptation to medium- to high-throughput screening settings are also highlighted.

Cardiotoxicity assessment using 3D vascularized cardiac tissue consisting of human iPSC-derived cardiomyocytes and fibroblasts

“…The discovery of induced pluripotent stem cells (iPSCs) and subsequent establishment of human iPSCderived cardiomyocytes (CMs) (hiPSC-CMs) have produced promising tools. 6 Using the hiPSC-CMs, development of an in vitro assay system to detect functional changes in CMs has been attempted. [7][8][9] The establishment of assay systems will enable accurate prediction of drug effects in humans without using experimental animals.…”

“…Various approaches to improve the maturity of hiPSC-CMs, including long-term culture, electrical pacing, and coculture with non-CMs, have been attempted. 18,19 Furthermore, the generation of three-dimensional (3D) cardiac tissues has also been developed, 6 and combination with different approaches was used to improve the maturity. 17,20 However, the method to generate matured cardiac tissues is complex and needs a long culture period.…”

Improving Generation of Cardiac Organoids from Human Pluripotent Stem Cells Using the Aurora Kinase Inhibitor ZM447439

“…HCOs are known to have a heart-like structure and cell composition [ 17 ]. They also have the advantage of being able to mimic the heart environment, allowing for accurate predictions of drug efficacy and safety evaluations, compared to conventional models [ 30 ]. There are two methods for producing hCOs using hiPSCs: (1) a direct method of differentiating each cell constituting the heart into two-dimensions (2D), followed by its conversion into a three-dimensional structure for co-culture [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 ]; and (2) a self-organizing method of creating a heart organ-like structure by aggregating stem cells [ 11 , 12 , 13 , 40 ].…”

Assessing Drug-Induced Mitochondrial Toxicity in Cardiomyocytes: Implications for Preclinical Cardiac Safety Evaluation

“…As for 3D cardiomyocyte models, hiPSC-CMs are the most common type of cardiomyocytes used in cardiac organoids. Other non-cardiomyocytes can be either induced from hiPSCs [332] or isolated from human tissue.…”