Tissue intermittent hypoxia (IH) occurs in obstructive sleep apnea, sickle cell anemia, physical
exercise and other conditions. Poor gas solubility and slow diffusion through culture media hampers
mimicking IH-induced transitions of O2 in vitro. We aimed to develop a system enabling exposure of
cultured cells to IH and to validate such exposure by real-time O2 measurements and cellular responses.
Standard 24-well culture plates and plates with bottoms made from a gas permeable film were placed
in a heated cabinet. Desired cycling of O2 levels was induced using programmable solenoids to purge
mixtures of 95% N2 + 5% CO2 or 95% O2 + 5% CO2. Dissolved oxygen, gas pressure, temperature, and
water evaporation were measured during cycling. IH-induced cellular effects were evaluated by hypoxia
inducible factor (HIF) and NF-κB luciferase reporters in HEK296 cells and by insulin secretion in rat
insulinoma cells. Oxygen cycling in the cabinet was translated into identical changes of O2 at the well
bottom in gas permeable, but not in standard cultureware. Twenty-four hours of IH exposure increased
HIF (112%), NF-κB (111%) and insulin secretion (44%). Described system enables reproducible and
prolonged IH exposure in cultured cells while controlling for important environmental factors.
Talk to us about your research requirements, and we’ll discuss customization options to ensure your Coy chamber meets your specifications.
