Advancements in molecular and bioengineering techniques for driving maturation of human pluripotent stem cell-derived cardiac three-dimensional models

This thesis describes essential steps required to enhance the utility and success of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) for drug screening and discovery applications, as well as for studying human cardiac development and disease. This thesis describes the successful generation of atrial-like hPSC-CMs using optimized protocols (chapter 2 and 3) and applied single cell transcriptomic analysis of subtype-specific cardiac models to decipher mechanisms governing their specification in vitro (chapter 7). Additionally, state-of-the art 3D culture methods, such as Engineered Heart Tissues (EHTs) (chapter 5) and their incorporation in a heart-on-chip microfluidic model (chapter 8) are developed to better recapitulate aspects of the complex physiology of the human heart. Subsequently, versatile analyses to further elucidate and mature the state of the hPSC-CMs are presented. For example, in chapter 8 and 9, different maturation strategies are used, such as the introduction of co-culture with different cardiac cell types and the development of a maturation media with a composition that resembles the one in the in vivo adult human heart. While there are still challenges to overcome and further progress to be made in the field of hPSC-CMs (chapter 4), this work represents a valuable contribution towards unlocking their full potential. This is shown in chapter 6, where cardiotoxic effects of anticancer drugs are evaluated as well as their long-term effect, being more clinically relevant. In summary, this thesis focuses entirely on the application and improvement of in vitro models for the heart, with stem cell technology at its core.