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Am J Physiol Lung Cell Mol Physiol 278: L858-L863, 2000;
1040-0605/00 $5.00
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Vol. 278, Issue 4, L858-L863, April 2000

SPECIAL COMMUNICATION
A method for measuring the oxygen consumption of intact cell monolayers

Kamel Mamchaoui and Georges Saumon

Institut National de la Santé et de la Recherche Médicale Unité 82, Faculté Xavier Bichat, 75018 Paris, France

This report describes an open-air method for measuring the O2 consumption (QO2) of intact monolayers of cultured cells. This method is based on Fick's second law of diffusion. It requires only a micromanipulator and a miniature O2 electrode to measure the PO2 gradient in the culture medium in the well. It was compared with the conventional oxygraph chamber method. Both methods gave the same value for QO2 in freshly isolated rat type II cells: 166 ± 15.3 nmol · h-1 · 106 cells-1 for the open-air method and 151 ± 11.6 nmol · h-1 · 106 cells-1 for the oxygraph chamber method (n = 11 experiments). But the open-air method gave significantly larger values for QO2 in cells cultured for 2 days (236 ± 8.8 nmol · h-1 · 106 cells-1) than the oxygraph method (71 ± 15.2 nmol · h-1 · 106 cells-1; P < 0.001; n = 12 experiments). This suggests that the way cells are detached from their substratum to be placed in the oxygraph chamber affects their QO2. The open-air method may be useful for studies on the metabolic properties of monolayers because the cells do not risk being damaged.

metabolism; type II pneumocytes; cell differentiation


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