AJP - Cell Physiology, Vol 265, Issue 5 C1266-C1270, Copyright © 1993 by American Physiological Society

ARTICLES

Microelectrode measurements of pericellular PO2 in erythropoietin-producing human hepatoma cell cultures

M. Wolff, J. Fandrey and W. Jelkmann
Department of Surgery, Rheinische Friedrich-Wilhelms-Universitat, Bonn, Germany.

On the basis of Fick's law of gas diffusion, it has been proposed that cells in conventional monolayer cultures may be severely hypoxic. Because knowledge of the cellular O2 availability is important for the interpretation of biochemical and toxicological cell culture work, microelectrode measurements of the pericellular PO2 were carried out using the erythropoietin (Epo)-producing human hepatoma cell lines Hep G2 and Hep 3B as an in vitro model. In confluent hepatoma cultures grown in polystyrene dishes and incubated in air with 5% CO2, the pericellular steady-state PO2 was < 1 mmHg. The rates of the production of immunoreactive Epo and lactate were high due to a misproportion between O2 supply and O2 requirements. Epo production decreased when shaken instead of static cultures were studied, or when the O2 concentration in the gas atmosphere was increased gradually up to 95%. In cultures grown on gas-permeable supports, pericellular and gas PO2 values were very similar, with increased Epo production at lowered PO2. In agreement with mathematical models, our experimental data make PO2 measurements desirable for studies of O2-dependent biological functions in cell cultures.

This article has been cited by other articles:

				J. Fandrey Oxygen-dependent and tissue-specific regulation of erythropoietin gene expression Am J Physiol Regulatory Integrative Comp Physiol, June 1, 2004; 286(6): R977 - R988. [Abstract] [Full Text] [PDF]

				C. B. Allen, B. K. Schneider, and C. W. White Limitations to oxygen diffusion and equilibration in in vitro cell exposure systems in hyperoxia and hypoxia Am J Physiol Lung Cell Mol Physiol, October 1, 2001; 281(4): L1021 - L1027. [Abstract] [Full Text] [PDF]

				K. Mamchaoui and G. Saumon A method for measuring the oxygen consumption of intact cell monolayers Am J Physiol Lung Cell Mol Physiol, April 1, 2000; 278(4): L858 - L863. [Abstract] [Full Text] [PDF]

				T. Hellwig-Burgel, K. Rutkowski, E. Metzen, J. Fandrey, and W. Jelkmann Interleukin-1beta and Tumor Necrosis Factor-alpha Stimulate DNA Binding of Hypoxia-Inducible Factor-1 Blood, September 1, 1999; 94(5): 1561 - 1567. [Abstract] [Full Text] [PDF]

				H. Mairbaurl, R. Wodopia, S. Eckes, S. Schulz, and P. Bartsch Impairment of cation transport in A549 cells and rat alveolar epithelial cells by hypoxia Am J Physiol Lung Cell Mol Physiol, October 1, 1997; 273(4): L797 - L806. [Abstract] [Full Text] [PDF]

				T. M. Casey, J. L. Pakay, M. Guppy, and P. G. Arthur Hypoxia Causes Downregulation of Protein and RNA Synthesis in Noncontracting Mammalian Cardiomyocytes Circ. Res., April 19, 2002; 90(7): 777 - 783. [Abstract] [Full Text] [PDF]