Open-volume oxygen consumption measurements for mammalian cell cultures

Abstract: Metabolism is currently a very relevant and popular area of research in cell biology. Studying metabolism cell respiration has to be taken into account. At the moment, the most used system is both expensive and requires the cells to be moved from their standard culturing conditions to other culture dishes. The different shape and plastic of these dishes could possibly influence the cells, and hence make it difficult to compare the data from these measurements to other measurements. In this study a system has been assembled and used to measure oxygen consumption under standard culturing conditions. The system is based on an article written by Mamchaoui et al. and uses the natural oxygen diffusion within the cell culture medium to measure the respiration. The oxygen pressure is measured at different depths within the cell culture medium, and when steady state is reached, Fick's laws are used to calculate the oxygen consumption. In order to measure the oxygen concentration at different depths, a system was assembled consisting of an oxygen measuring system with fiber optic microsensors, a motorized vertical stage and a rigid table top pole stand. The system was evaluated by testing the reproducability of measurements, and it showed standard deviations comparable to those acquired with another system. To conclude, it was shown that the system could be produced with less than one fifth of the cost of the Seahorse system, it can measure under normal culturing conditions and shows measuring errors close to those acquired with established measuring systems.