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Glucose production assay

LW Lingdi Wang
IS Iain Scott
LZ Lu Zhu
KW Kaiyuan Wu
KH Kim Han
YC Yong Chen
MG Marjan Gucek
MS Michael N. Sack
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Glucose production from primary hepatocytes was measured using a colorimetric glucose oxidase assay (Sigma). Briefly, primary hepatocytes were isolated and cultured in DMEM medium containing 10% FBS for 24 h, and then were cultured in glucose production medium (glucose-free DMEM, pH 7.4, containing 20 mM sodium lactate and 2 mM sodium pyruvate, without phenol red) overnight to deplete glycogen, followed by incubated for another 5 h at 37 °C, 5% CO2, in glucose production buffer with or without 100 nM glucagon. The medium was collected for glucose measurement. The glucose assays were conducted in triplicate.

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