In vitro enzymatic electrochemical monitoring of glucose metabolism and production in rat primary hepatocytes on highly O2 permeable plates

Abstract

In situ continuous glucose monitoring under physiological culture conditions is imperative in understanding the dynamics of cell and tissue behaviors and their physiological responses since glucose plays an important role in principal source of biological energy. We therefore examined physiologically relevant dynamic changes in glucose levels based on glucose metabolism and production during aerobic culture (10% O2) of rat primary hepatocytes stimulated with insulin or glucagon on a highly O2 permeable plate, which can maintain the oxygen concentration close to the periportal zone of the liver. As glucose monitoring devices, we used oxygen-independent glucose dehydrogenase-modified single-walled carbon nanotube electrodes placed close to the surface of the hepatocytes. The current response of glucose oxidation slightly decreased after the addition of insulin in the presence of glucose due to the acceleration of glucose uptake by the hepatocytes, whereas that significantly increased after the addition of glucagon and fructose even in the absence of glucose due to the conversion of fructose to glucose based on gluconeogenesis. These phenomena might be consistent relatively with the physiological behaviors of hepatocytes in the periportal region. The present monitoring system would be useful for the studies of glucose homeostasis and diabetes in vitro.