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Evaluation of glucose transport and its regulation by insulin in human monocytes using flow cytometry
Author(s) -
Dimitriadis George,
Maratou Eirini,
Boutati Eleni,
Psarra Katherina,
Papasteriades Chryssa,
Raptis Sotirios A.
Publication year - 2005
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20108
Subject(s) - glut3 , glucose transporter , glut4 , glut1 , glucose uptake , insulin , flow cytometry , monocyte , medicine , cd14 , glucose transporter type 1 , endocrinology , chemistry , biology , microbiology and biotechnology
Background We investigated the effects of insulin on glucose transport in human monocytes using flow cytometry, a method with several advantages over previously used techniques. We hypothesized that monocytes could be used as tools to study insulin action at the cellular level and facilitate the investigation of mechanisms that lead to insulin resistance. Methods Blood was withdrawn from 38 healthy subjects. The expression of glucose transporter (GLUT) isoforms in plasma membrane and the rates of glucose transport were determined with and without insulin (10 to 1,000 mU/L). Anti‐CD14 phycoerythrin monoclonal antibody was used for monocyte gating. GLUT isoforms were determined after staining cells with specific antisera to GLUT1, GLUT3, and GLUT4. Glucose transport was monitored with 6‐[ N ‐(7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)amino]‐6‐deoxyglucose (NBDG). Results Insulin increased the uptake of NBDG (median effective dose 20 mU/L) and the expression of GLUT3 and GLUT4 isoforms in the plasma membrane (median effective doses 20 and 35 mU/L, respectively) but had no effect on GLUT1. Maximal effects were always reached at 100 mU/L of insulin. Conclusions Monocytes may be a valid model system to study the effects of insulin on glucose transport. Further, flow cytometry is suitable for this investigation and can be used as an alternative to radiotracer methods. © 2005 Wiley‐Liss, Inc.