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Metabolism / Mitochondria

Aerobic metabolism is an efficient pathway for ATP production in mammalian cells. However, ‘standard’ cell culture methods include non-physiological exposures to both antibiotics and also room-air levels of oxygen. While convenient for preventing bacterial contamination, use of antibiotics in cell culture induces mitochondrial dysfunction and oxidative damage. While physiologic oxygen (physioxia) for most tissues is <5%, ‘normal’ exposure of room-air oxygen (21% O2) results in ROS artifact, altered cell function, and questionably relevant in vitro data.

Antibiotic-free, Physioxic conditions for cell culture:

  • reduce ROS artifact
  • facilitate metabolic reprogramming
  • regulate immunometabolism
  • improve mitochondrial morphology
  • modulate metabolic flux
  • control metabolic zonation
  • maintain the metabolome
  • reduce mitochondrial stress
New tools allow researchers to aseptically perform cell-based studies without antibiotics under physiologically relevant O2 conditions during all steps of cell incubation, cell handling, and cell analysis. Incubator subchambers upgrade the function of existing CO2 incubators. Cell-culture workstations provide full-time protection from contamination & room-air oxygen during the incubation, handling and analysis of cell cultures.

Cell Equipment

ProOx C21 & C-Chamber

Cell Research – physiologic oxygen chamber, hypoxia chamber with static control of O2 and CO2ProOx C21 & C-Chamber

Xvivo System

Cell Research – aseptic, physiologic oxygen incubator and hood system, hypoxia workstation, hypoxia glove box with independently programmable control of O2, CO2, and Temperature in one (or more) modular incubator chamber(s) and/or cell handling chamber(s) – Xvivo System

Discover More


He, Yang-Yang et al. Identification of Hypoxia Induced Metabolism Associated Genes in Pulmonary Hypertension. Front. Pharmacol 2021: 12:753727. doi: 10.3389/fphar.2021.753727

Surace, L. et al. Dichotomous metabolic networks govern human ILC2 proliferation and function. Nat Immunol. 2021; 22, 1367–1374 . doi: 10.1038/s41590-021-01043-8

Elliott RL, Jiang XP. The adverse effect of gentamicin on cell metabolism in three cultured mammary cell lines: “Are cell culture data skewed?”. PLoS One. 2019;14(4):e0214586. Published 2019 Apr 1. doi:10.1371/ journal.pone. 0214586. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6443165/

Timpano S., et al., Physioxic human cell culture improves viability, metabolism, and mitochondrial morphology while reducing DNA damage. FASEB J. 2019 Apr;33(4):5716-5728. https://www.ncbi.nlm.nih.gov/pubmed/30649960

Korski KI., et al., Hypoxia Prevents Mitochondrial Dysfunction and Senescence in Human c-Kit(+) Cardiac Progenitor Cells. Stem Cells. 2019 Apr;37(4):555-567. https://www.ncbi.nlm.nih.gov/pubmed/30629785

Stuart JA., et al., How Supraphysiological Oxygen Levels in Standard Cell Culture Affect Oxygen-Consuming Reactions., Oxid Med Cell Longev., 2018 Sep 30;2018:8238459. https://www.ncbi.nlm.nih.gov/pubmed/30363917

Jaber SM., et al., Sex differences in the mitochondrial bioenergetics of astrocytes but not microglia at a physiologically relevant brain oxygen tension. Neurochem Int. 2018 Jul;117:82-90. https://www.ncbi.nlm.nih.gov/pubmed/28888963


Browse Metabolism Research publications citing equipment from BioSpherix Ltd


Brochure – Cell Based Assays Literature – Seahorse

Brochure – Tips for Planning, Executing and Reporting In Vitro studies at Physiologically Relevant Oxygen Levels

Blog Posts

Cytocentric Blog – Interview with Cytocentric Visionary: Jeffrey A. Stuart, Brock University. Oxygen: A Big Uncontrolled Variable in Cell Culture

Lab References

Mitochondrial Research at McGill University

Metabolic Research at Tufts Medical Center

Metabolic Research at RESTORE (Toulouse)