Method for measuring a comprehensive energy budget in a proliferating cell system over multiple cell cycles

Isolated cell systems are now being used very effectively to study a range of important biochemical questions, but their energy metabolism has never been comprehensively investigated. We have developed a system, using J2E cells, which enables us to measure total ATP turnover and the contribution of various fuels and pathways to this total in a dynamic, proliferating preparation. Cells are cultured in 500 mi airtight glass containers which enables (1) the measurement of oxygen consumption, (2) the collection and measurement of (CO2)-C-14 production from labelled fuels, and (3) the measurement of metabolite utilization and production. Data on cell numbers are then used to produce a curve of cell number vs. time, the area under which (cell numbers hour) is used as a base by which all measurements and experiments are compared. To our knowledge this is the first time a comprehensive energy budget has been measured in a proliferating cell system over a period that covers multiple cell cycles. (C) 1997 Wiley-Liss, Inc.