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Novel Method for the Detection of Glucose Uptake: Direct Measurement of Glucose Levels in Cultured Cells
Author(s) -
Yan Kevin,
Li Shunan,
Lin Yuan,
Saddar Sonika,
Tchaga Grigoriy,
Yan Gordon
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.627.5
Subject(s) - glucose oxidase , glucose transporter , carbohydrate metabolism , glucose uptake , biochemistry , metabolism , cell culture , enzyme , chemistry , snf3 , biology , endocrinology , insulin , genetics
Glucose uptake is the transport process of glucose from the outside of cells into cells across the cell membrane. It is one of the key regulation steps of glucose metabolism. Study of glucose uptake provides important information for understanding glucose metabolism and its regulation in normal and disease development. Here we report the development of a novel glucose uptake detection method based on direct measurement of glucose levels in cultured cells. In this method, we utilized the inhibitor, 3‐bromopyruvate (3BP), to temporarily inhibit hexokinases, the first enzyme that metabolizes glucose in cells. Thus, glucose metabolism was blocked, which leads to glucose accumulation rather than its rapid metabolism inside the cells. Glucose uptake was then detected by direct measurement of the glucose levels inside of the cells using Glucose Oxidase/HRP/OxiRed TM enzyme based method. In comparison with current widely used methods, this direct glucose uptake detection method has the following advantages: i) No radioisotope materials are used; ii) It directly detects glucose without using structurally modified glucose derivatives; iii) Glucose from culture medium is transported inside of cells, and then detected with minimum medium changes and washing processes. This simple, sensitive and direct glucose uptake detection method provides a powerful tool for studying the process of glucose uptake and its regulation, as well as for screening and characterization of drugs that regulate glucose uptake during normal and disease development. Support or Funding Information This project was supported by the New Product Development Funds from R&D Division of BioVision Inc.