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

Barreto, S; Al-Zubaidi, MA; Dale, TP; Worrall, AJ; Kapacee, Z; Kimber, SJ; Sulé-Suso, J; Forsyth, NR; Rutter, AV

Abstract

<jats:p>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.</jats:p>

Citation

Barreto, S., Al-Zubaidi, M., Dale, T., Worrall, A., Kapacee, Z., Kimber, S., …Rutter, A. (2022). Physiological Oxygen Causes the Release of Volatile Organic Compounds from Human Pluripotent Stem Cells with Possible Roles in Maintaining Self-Renewal and Pluripotency. Preprints, https://doi.org/10.20944/preprints202203.0143.v1

• Journal Article Type: Article
• Acceptance Date: Mar 10, 2022
• Publication Date: Mar 10, 2022
• Journal: Preprints
• Publisher: MDPI
• Peer Reviewed: Not Peer Reviewed
• DOI: https://doi.org/10.20944/preprints202203.0143.v1
• Publisher URL: https://www.preprints.org/manuscript/202203.0143/v1