Dissolved oxygen and its availability to cells in culture is an overlooked variable which can have significant consequences on experimental research outcomes, including reproducibility. Oxygen sensing pathways play key roles in cell growth and behavior and pericellular oxygen levels should be controlled when establishing models. Standard cell culture techniques do not have adequate control over pericellular oxygen levels. Slow diffusion through culture media limits the precision of oxygen delivery to cells, making it difficult to accurately reproduce -like oxygen conditions. Furthermore, different types of cells consume oxygen at varying rates and this can be affected by the density of growing cells. Here, we describe a novel system that utilizes hypoxic chambers and oxygen-permeable culture dishes to control pericellular oxygen levels and provide rapid oxygen delivery to adherent cells. This procedure is particularly relevant for protocols studying effects of rapid oxygen changes or intermittent hypoxia on cellular behavior. The system is inexpensive and easily assembled without highly specialized equipment.
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http://dx.doi.org/10.21769/BioProtoc.3371 | DOI Listing |
Free Radic Biol Med
January 2025
Conway Institute of Biomolecular and Biomedical Research and School of Medicine, University College Dublin, Belfield, Dublin, Ireland. Electronic address:
Oxygen (O)-controlled cell culture has been pivotal in studying mammalian mechanisms of O sensing, regulation, and utilization. We posit, however, that O-controlled cell culture is paradoxically not controlling O. There is overwhelming evidence that the pericellular O is lower than the surrounding gas phase due to cellular O consumption.
View Article and Find Full Text PDFMethods Mol Biol
November 2024
CNC-UC, Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.
In vitro and ex vivo studies are crucial for mitochondrial research, offering valuable insights into cellular mechanisms and aiding in diagnostic and therapeutic strategies. Accurate in vitro models rely on adequate cell culture conditions, such as the composition of culture media and oxygenation levels. These conditions can influence energy metabolism and mitochondrial activities, thus impacting studies involving mitochondrial components, such as the effectiveness of anticancer drugs.
View Article and Find Full Text PDFCell Host Microbe
September 2024
State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, P.R. China; Key Laboratory of Biopharmaceutical Preparation and Delivery, Chinese Academy of Sciences, Beijing 100190, P.R. China; School of Chemical Engineering, University of Chinese Academy of Sciences, Beijing 100049, P.R. China. Electronic address:
Pflugers Arch
September 2024
Department of Biological Sciences, Brock University, St. Catharines, ON, L2S 3A1, Canada.
Cellular responses to hypoxia are crucial in various physiological and pathophysiological contexts and have thus been extensively studied. This has led to a comprehensive understanding of the transcriptional response to hypoxia, which is regulated by hypoxia-inducible factors (HIFs). However, the detailed molecular mechanisms of HIF regulation in hypoxia remain incompletely understood.
View Article and Find Full Text PDFFEBS Open Bio
August 2024
Department of Diagnostic Radiology and Nuclear Medicine, University of Maryland School of Medicine, Baltimore, MD, USA.
In multicellular organisms, stem cells are impacted by microenvironmental resources such as nutrient availability and oxygen tension for their survival, growth, and differentiation. However, the accessibility of these resources in the pericellular environment greatly varies from organ to organ. This divergence in resource availability leads to variations in the potency and differentiation potential of stem cells.
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