(F1078) CONTINUOUS PHYSIOLOGICAL OXYGEN INCREASES ATRIAL RATHER THAN VENTRICULAR SPECIFICATION OF HUMAN INDUCED PLURIPOTENT STEM CELL DERIVED CARDIOMYOCYTES
Professor/PI Odense University Hospital/University of Southern Denmark, Denmark
Abstract: The human heart is unable to replace cardiomyocytes (CMs) lost following myocardial infarction, a major cause of death worldwide. In standard care, curative treatments for MI lack, but recent advances in generating CMs from induced pluripotent stem cells (iPSC-CMs) has renewed hope for a feasible therapy of MI patients. Despite this progress, current iPSC-CMs fail to fully reproduce the physiology of primary adult CMs, and one contributor might be a sub-optimal laboratory environment, including exposure to atmospheric O2 tension rather than the much lower physiological O2 tension experienced by cardiac stem cells during cardiac development. At present, we investigated the effect of physiological O2 tension on CM derivation as compared to iPSC-CM derived under atmospheric O2. For manufacturing purposes, we exploited a highly specialized GMP-compliant quad SCI-TIVE glovebox compartment with sensitive surveillance, enabling accurate monitoring of gas composition and temperature to ensure continuous and stable specified O2 tensions. We found that iPSC-CMs derived under theoretical physiological O2 levels displayed less ventricular specification, higher ploidy, and a lower binucleation rate as compared to iPSC-CMs derived at atmospheric O2. Single cell transcriptomics (scRNAseq) showed that iPSC-CMs at physiological O2 were more atrial-like, and this was confirmed by comparing to large scRNAseq datasets of atrial and ventricular CMs. Finally, in vitro functionality studies showed that iPSC-CMs at physiological low O2 exhibited less beating and calcium waves further supportive of an atrial phenotype. In perspectives, these data provide new biological insights essential for manufacturing of iPSC-CM cell products that may be used for regenerative or diagnostic purposes of the heart.
Funding Source: The Novo Nordisk Foundation (#NNF17OC0028764; #NNF19OC0055353) Danish Cardiovascular Academy (#PD2Y-2021004-DCA-67242) Danish Independent Research (# 8045-00019B and #4285-00027B) The Lundbeck Foundation (#R313-2019-573)
