METHOD FOR MEASURING A COMPREHENSIVE ENERGY BUDGET IN A PROLIFERATINGCELL SYSTEM OVER MULTIPLE CELL-CYCLES

Isolated cell systems are now being used very effectively to study a r ange of important biochemical questions, but their energy metabolism h as 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 dyna mic, proliferating preparation. Cells are cultured in 500 mi airtight glass containers which enables (1) the measurement of oxygen consumpti on, (2) the collection and measurement of (CO2)-C-14 production from l abelled fuels, and (3) the measurement of metabolite utilization and p roduction. Data on cell numbers are then used to produce a curve of ce ll number vs. time, the area under which (cell numbers hour) is used a s a base by which all measurements and experiments are compared. To ou r knowledge this is the first time a comprehensive energy budget has b een measured in a proliferating cell system over a period that covers multiple cell cycles. (C) 1997 Wiley-Liss, Inc.