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Online Measurement of Glucose Consumption from HepG2 Cells Using an Integrated Bioreactor and Enzymatic Assay

Author:
Adams, Anna G., Bulusu, Radha Krishna Murthy, Mukhitov, Nikita, Mendoza-Cortes, Jose L., Roper, Michael G.
Source:
Analytical chemistry 2019 v.91 no.8 pp. 5184-5190
ISSN:
1520-6882
Subject:
bioreactors, fluorescence, glucose, hepatocytes, hepatoma, homeostasis, human cell lines, humans, insulin, ketone bodies, lactic acid, liver, metabolism, mixing, pancreas, pyruvic acid
Abstract:
Hepatocytes help to maintain glucose homeostasis in response to a variety of signals, including pancreatic hormones such as insulin. Insulin is released from the pancreas with variable dynamics, yet the role that these play in regulating glucose metabolism in the liver is still unclear. In this study, a modular microfluidic system was developed to quantitatively measure the effect of insulin dynamics on glucose consumption by a human hepatocarcinoma cell line, HepG2. A microfluidic bioreactor that contained 10⁶ HepG2 cells was cultured for up to 10 days in an incubator. For glucose consumption experiments, the bioreactor was removed from the incubator and connected with reagents for an enzymatic glucose assay. The mixed components were then delivered into a droplet-based microfluidic system where the intensity of the fluorescent product of the enzyme assay was used to quantify the glucose concentration. By optimizing the mixing time of the reagents, the dynamic range of the enzymatic assay was adjusted to 0–12 mM glucose and had a time resolution of 96 ± 12 s. The system was used to observe rapid changes in insulin-induced glucose consumption from HepG2 cells. This assay format is versatile and can be expanded to measure a variety of hepatic metabolites, such as lactate, pyruvate, or ketone bodies, which will enable the correlation of pancreatic hormone dynamics to liver metabolism.
Agid:
6371899