Introduction

‍Cardiac tissue engineering strategies designed to repair the heart following myocardial infarction (MI), along with projects aimed at understanding how coronary microvascular disease (CMD) can lead to the development of heart failure (HF), require well-characterised and stable coronary microvascular endothelial cells that can be generated at scale without using xenogeneic materials. This study outlines the development and characterisation of phenotypically stable human pluripotent stem cell-derived cardiac microvascular-like endothelial cells (hPSC-CMVECs) and cardiac pericyte-like cells for such endeavours.

Methods

‍hPSCs were subjected to a growth factor differentiation protocol within 3D stirred tank bioreactors, resulting in the generation of vascular organoids. hPSC-ECs emerging from the dissociation of these organoids underwent high concentration VEGF-A treatment and were examined via single cell-RNA sequencing (scRNA-seq), alongside the non-endothelial component of the organoid.

Results

‍The VEGF-A-treated hPSC-ECs were phenotypically stable, and scRNA-seq revealed robust expression of cardiac endothelial- and microvascular-associated genes. Further, these cells clustered closely with bona fide cardiac microvascular endothelial cells isolated from human hearts. scRNA-seq analysis of the non-endothelial component of the vascular organoid highlighted a gene expression profile indicative of cardiac pericytes.

Conclusion

‍The scalable and xenogeneic-free 3D protocol described here allows for the production of hPSC-CMVECs with high angiogenic potential and cardiac pericyte-like cells. The absence of Matrigel enhances the translational capacity of these cells that could be evaluated in pre-clinical models of post-MI cardiac regeneration or utilised for the in vitro investigation of the CMD-HF axis.

‍