Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Physiological Oxygen Causes the Release of Volatile Organic Compounds from Human Pluripotent Stem Cells with Possible Roles in Maintaining Self-Renewal and Pluripotency

Version 1 : Received: 3 March 2022 / Approved: 10 March 2022 / Online: 10 March 2022 (10:23:46 CET)

How to cite: Barreto, S.; Al-Zubaidi, M.A.; Dale, T.P.; Worrall, A.J.; Kapacee, Z.; Kimber, S.J.; Sulé-Suso, J.; Forsyth, N.R.; Rutter, A.V. Physiological Oxygen Causes the Release of Volatile Organic Compounds from Human Pluripotent Stem Cells with Possible Roles in Maintaining Self-Renewal and Pluripotency. Preprints 2022, 2022030143 (doi: 10.20944/preprints202203.0143.v1). Barreto, S.; Al-Zubaidi, M.A.; Dale, T.P.; Worrall, A.J.; Kapacee, Z.; Kimber, S.J.; Sulé-Suso, J.; Forsyth, N.R.; Rutter, A.V. Physiological Oxygen Causes the Release of Volatile Organic Compounds from Human Pluripotent Stem Cells with Possible Roles in Maintaining Self-Renewal and Pluripotency. Preprints 2022, 2022030143 (doi: 10.20944/preprints202203.0143.v1).

Abstract

Human pluripotent stem cells (hPSCs) have widespread potential biomedical applications. There is a need for large-scale in vitro production of hPSCs, and optimal culture methods are vital in achieving this. Physiological oxygen (2% O2) improves key hPSCs attributes, including genomic integrity, viability, and clonogenicity, however, its impact on hPSC metabolism remains un-clear. Here, Selected Ion Flow Tube-Mass Spectrometry (SIFT-MS) was used to detect and quantify metabolic Volatile Organic Compounds (VOCs) in the headspace of hPSCs and their differentiated progeny. hPSCs were cultured in either 2% O2 or 21% O2. Media was collected from cell cultures and transferred into glass bottles for SIFT-MS measurement. The VOCs acetaldehyde and dimethyl sulfide (DMS)/ethanethiol were significantly increased in undifferentiated hPSCs compared to their differentiating counterparts, and these observations were more apparent in 2% O2. Pluripotent marker expression was consistent across both O2 concentrations tested. Transcript levels of ADH4, ADH5, and CYP2E1, encoding enzymes involved in converting ethanol to acetaldehyde, were upregulated in 2% O2, and chemical inhibition of ADH and CYP2E1 decreased acetaldehyde levels in hPSCs. Acetaldehyde and DMS/ethanethiol may be indicators of altered metabolism pathways in physiological oxygen culture conditions. The identification of non-destructive biomarkers for hPSC characterization has the potential to facilitate large-scale in vitro manufacture for future biomedical application.

Keywords

human pluripotent stem cells; human embryonic stem cells; human induced pluripotent stem cells; cellular metabolism; oxygen; self-renewal; pluripotency; differentiation; volatile organic compounds; Selected ion flow tube-mass spectrometry

Subject

LIFE SCIENCES, Cell & Developmental Biology

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