PT  - JOURNAL ARTICLE
AU  - Joycelyn Tan
AU  - Sam Virtue
AU  - Dougall M. Norris
AU  - Olivia J. Conway
AU  - Ming Yang
AU  - Christopher Gribben
AU  - Fatima Lugtu
AU  - James R. Krycer
AU  - Richard J. Mills
AU  - Ioannis Kamzolas
AU  - Conceição Pereira
AU  - Martin Dale
AU  - Amber S. Shun-Shion
AU  - Harry J. M. Baird
AU  - James A. Horscroft
AU  - Alice P. Sowton
AU  - Marcella Ma
AU  - Stefania Carobbio
AU  - Evangelia Petsalaki
AU  - Andrew J. Murray
AU  - David C. Gershlick
AU  - James E. Hudson
AU  - Ludovic Vallier
AU  - Kelsey H. Fisher-Wellman
AU  - Christian Frezza
AU  - Antonio Vidal-Puig
AU  - Daniel J. Fazakerley
TI  - Oxygen is a critical regulator of cellular metabolism and function in cell culture
AID  - 10.1101/2022.11.29.516437
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.11.29.516437
4099  - http://biorxiv.org/content/early/2022/11/29/2022.11.29.516437.short
4100  - http://biorxiv.org/content/early/2022/11/29/2022.11.29.516437.full
AB  - Cell culture, the workhorse of biomedical research, is generally considered to be hyperoxic. However, oxygen consumption by cells is underappreciated. High cellular respiration rates can rapidly deplete oxygen, resulting in local hypoxia. Increasing pericellular oxygen levels rewired the metabolism of multiple post-mitotic cell-lines, both in monolayer and organoid culture. Under standard conditions, cultured adipocytes are hypoxic and highly glycolytic. Increased oxygen availability diverted glucose flux toward mitochondria and increased lipogenesis from glucose-derived carbon. These metabolic changes were coupled to thousands of gene expression changes, and rendered adipocytes more sensitive to insulin and lipolytic stimuli. Importantly, pathway analyses revealed increasing oxygen tension made in vitro adipocytes more similar to in vivo adipose tissue. hPSC-derived hepatocytes and cardiac organoids were also functionally enhanced by increased oxygen. Our findings suggest that oxygen is limiting in many standard cell culture systems, and highlight how controlling oxygen availability can improve translatability of cell models.Competing Interest StatementThe authors have declared no competing interest.