Glucose Dependency of the Metabolic Pathway of HEK 293 Cells Measured by a Flow-through Type pH/CO₂ Sensor System Using ISFETs

抄録

Our group previously reported the application of a flow-through type pH/CO₂ sensor system designed to evaluate the metabolic activity of cultured cells. The sensor system consists of two ion-sensitive field effect transistors (ISFETs), an ISFET to measure the total pH change and an ISFET enclosed within a gas-permeable silicone tube to measure the pH change attributable to CO₂. In that study, we used the system to quantitatively analyze metabolic switching induced by glucose concentration changes in three cultured cell types (bovine arterial endothelium cell (BAEC), human umbilical vein endothelium cell (HUVEC), and rat cardiomuscle cell (RCMC)), and to measure the production rates of total carbonate and free lactic acid in the cultured cells. In every cell type examined, a decrease in the glucose concentration led to an increase in total carbonate, a product of cellular respiration, and a decrease of free lactic acid, a product of glycolysis. There were very significant differences among the cell types, however, in the glucose concentrations at the metabolic switching points. We postulated that the cell has a unique switching point on the metabolic pathway from glycolysis to respiration. In this paper we use our sensor system to evaluate the metabolic switching of human embryonic kidney 293 cells triggered by glucose concentration changes. The superior metabolic pathway switched from glycolysis to respiration when the glucose concentration decreased to about 2 mM. This result was very similar to that obtained in our earlier experiments on HUVECs, but far different from our results on the other two cells types, BAECs and RCMCs. This sensor system will be useful for analyzing cellular metabolism for many applications and will yield novel information on different cell types.