An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistance

Tumor necrosis factor alpha (TNF-α)/hypoxia-treated 3T3-L1 adipocytes have been used to model inflamed and insulin-resistant adipose tissue: this study examines gaps in the model. We tested whether modulating TNF-α/hypoxia treatment time could reduce cell death while still inducing inflammation and...

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Main Authors: Ifeoluwa A. Odeniyi, Bulbul Ahmed, Benjamin Anbiah, Grace Hester, Peter T. Abraham, Elizabeth A. Lipke, Michael W. Greene
Format: Article
Language:English
Published: Taylor & Francis Group 2024-12-01
Series:Adipocyte
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Online Access:https://www.tandfonline.com/doi/10.1080/21623945.2024.2414919
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author Ifeoluwa A. Odeniyi
Bulbul Ahmed
Benjamin Anbiah
Grace Hester
Peter T. Abraham
Elizabeth A. Lipke
Michael W. Greene
author_facet Ifeoluwa A. Odeniyi
Bulbul Ahmed
Benjamin Anbiah
Grace Hester
Peter T. Abraham
Elizabeth A. Lipke
Michael W. Greene
author_sort Ifeoluwa A. Odeniyi
collection DOAJ
description Tumor necrosis factor alpha (TNF-α)/hypoxia-treated 3T3-L1 adipocytes have been used to model inflamed and insulin-resistant adipose tissue: this study examines gaps in the model. We tested whether modulating TNF-α/hypoxia treatment time could reduce cell death while still inducing inflammation and insulin resistance. Adipocytes were treated with TNF-α (12 h or 24 h) and incubated in a hypoxic chamber for 24 h. To examine maintenance of the phenotype over time, glucose and FBS were added at 24 h post initiation of treatment, and the cells were maintained for an additional 48 h. Untreated adipocytes were used as a control. Viability, insulin resistance, and inflammation were assessed using Live/Dead staining, RT-qPCR, ELISA, and glucose uptake assays. Treatment for 12 h with TNF-α in the presence of hypoxia resulted in an increase in the percentage of live cells compared to 24 h treated cells. Importantly, insulin resistance and inflammation were still induced in the 12 h treated adipocytes: the expression of the insulin sensitive and inflammatory genes was decreased and increased, respectively. In 72 h treated adipocytes, no significant differences were found in the viability, glucose uptake or insulin-sensitive and inflammatory gene expression. This study provides a modified approach to in vitro odeling adipocyte inflammation and insulin resistance.
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spelling doaj-art-fc2de475e9e44ab38af824874cbf36c12024-12-09T07:23:12ZengTaylor & Francis GroupAdipocyte2162-39452162-397X2024-12-0113110.1080/21623945.2024.2414919An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistanceIfeoluwa A. Odeniyi0Bulbul Ahmed1Benjamin Anbiah2Grace Hester3Peter T. Abraham4Elizabeth A. Lipke5Michael W. Greene6Department of Nutritional Sciences, Auburn University, Auburn, AL, USADepartment of Nutritional Sciences, Auburn University, Auburn, AL, USADepartment of Chemical Engineering, Auburn University, Auburn, AL, USADepartment of Chemical Engineering, Auburn University, Auburn, AL, USADepartment of Chemical Engineering, Auburn University, Auburn, AL, USADepartment of Chemical Engineering, Auburn University, Auburn, AL, USADepartment of Nutritional Sciences, Auburn University, Auburn, AL, USATumor necrosis factor alpha (TNF-α)/hypoxia-treated 3T3-L1 adipocytes have been used to model inflamed and insulin-resistant adipose tissue: this study examines gaps in the model. We tested whether modulating TNF-α/hypoxia treatment time could reduce cell death while still inducing inflammation and insulin resistance. Adipocytes were treated with TNF-α (12 h or 24 h) and incubated in a hypoxic chamber for 24 h. To examine maintenance of the phenotype over time, glucose and FBS were added at 24 h post initiation of treatment, and the cells were maintained for an additional 48 h. Untreated adipocytes were used as a control. Viability, insulin resistance, and inflammation were assessed using Live/Dead staining, RT-qPCR, ELISA, and glucose uptake assays. Treatment for 12 h with TNF-α in the presence of hypoxia resulted in an increase in the percentage of live cells compared to 24 h treated cells. Importantly, insulin resistance and inflammation were still induced in the 12 h treated adipocytes: the expression of the insulin sensitive and inflammatory genes was decreased and increased, respectively. In 72 h treated adipocytes, no significant differences were found in the viability, glucose uptake or insulin-sensitive and inflammatory gene expression. This study provides a modified approach to in vitro odeling adipocyte inflammation and insulin resistance.https://www.tandfonline.com/doi/10.1080/21623945.2024.2414919Adipocyteinflammationinsulin resistancein vitro model3T3-L1 cells
spellingShingle Ifeoluwa A. Odeniyi
Bulbul Ahmed
Benjamin Anbiah
Grace Hester
Peter T. Abraham
Elizabeth A. Lipke
Michael W. Greene
An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistance
Adipocyte
Adipocyte
inflammation
insulin resistance
in vitro model
3T3-L1 cells
title An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistance
title_full An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistance
title_fullStr An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistance
title_full_unstemmed An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistance
title_short An improved in vitro 3T3-L1 adipocyte model of inflammation and insulin resistance
title_sort improved in vitro 3t3 l1 adipocyte model of inflammation and insulin resistance
topic Adipocyte
inflammation
insulin resistance
in vitro model
3T3-L1 cells
url https://www.tandfonline.com/doi/10.1080/21623945.2024.2414919
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