Home General Physiology and Biophysics 2015 General Physiology and Biophysics Vol.34, No.3, p.235–247, 2015

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Quarterly, 80 pp. per issue
Founded: 1982
ISSN  1338-4325 (online)

Published in English

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General Physiology and Biophysics Vol.34, No.3, p.235–247, 2015

Title: System for exposing cultured cells to intermittent hypoxia utilizing gas permeable cultureware
Author: Jan Polak, Karen Studer-Rabeler, Holly McHugh, Mehboob A. Hussain, Larissa A. Shimoda

Abstract: 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.

Keywords: Oxygen — Cell culture — Insulin secretion — Hypoxia inducible factor 1 — NF-κB
Year: 2015, Volume: 34, Issue: 3 Page From: 235, Page To: 247

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