Dogan, F, Al Jumaily, RK, Kitchen, M and Forsyth, N (2022) Physoxia influences global and gene-specific methylation in pluripotent stem cells. bioRxiv.

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Pluripotent stem cells (PSC) possess unlimited proliferation, self-renewal, and a differentiation capacity spanning all germ layers. Appropriate culture conditions are important for the maintenance of self-renewal, pluripotency, proliferation, differentiation, and epigenetic states. Oxygen concentrations vary across different human tissues depending on precise cell location and proximity to vascularisation. The bulk of PSC culture-based research is performed in a physiologically hyperoxic, air oxygen (21% O2) environment, with numerous reports now detailing the impact of physiologic normoxia (physoxia) low oxygen culture in the maintenance of stemness, survival, morphology, proliferation, differentiation potential, and epigenetic profiles. Epigenetic mechanisms affect multiple cellular characteristics including gene expression during development and cell fate determination in differentiated cells. We hypothesized that epigenetic marks are responsive to a reduced oxygen microenvironment in PSCs and their differentiation progeny. Here, we evaluated the role of physoxia in PSC culture, the regulation of DNA methylation (5mC and 5hmC), and expression of regulatory enzymes DNMTs and TETs. Physoxia enhanced the functional profile of PSC including proliferation, metabolic activity, and stemness attributes. PSCs cultured in physoxia revealed significant downregulation of DNMT3B, DNMT3L, TET1, and TET3 vs. air oxygen, accompanied by significantly reduced 5mC and 5hmC levels. Downregulation of DNMT3B was associated with an increase in its promoter methylation. Coupled to above we also noted decreased HIF1A but increased HIF2A expression in physoxia cultured PSCs, versus air oxygen. In conclusion, PSCs display oxygen-sensitive methylation patterns that correlate with transcriptional and translational regulation of the de novo methylase DNMT3B.

Item Type: Article
Additional Information: This preprint article has now been published as Gold Open Access via MDPI at International Journal of Molecular Sciences journal, where information can be found in a separate EPrints ID. For any relevant information, please refer to the publisher version or bioRxiv. The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (
Subjects: R Medicine > R Medicine (General)
Divisions: Faculty of Medicine and Health Sciences > School of Pharmacy and Bioengineering
Related URLs:
Depositing User: Symplectic
Date Deposited: 16 May 2023 14:24
Last Modified: 16 May 2023 14:24

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