Event Title

The Effects of Caffeine on Insulin Resistant 3T3-L1 Adipocytes

Presenter Information

Jillian Batten
Kristen Kaytes

Faculty Advisor

Dr. Tom Peeler

Start Date

24-4-2018 4:00 PM

End Date

24-4-2018 5:00 PM

Description

The 3T3-L1 cell line is a commonly used model system in studies on obesity and adipogenesis due to the ease and speed that 3T3-L1 fibroblasts can be differentiated into mature adipocytes. Previous studies have shown that insulin resistance can be induced in mature 3T3-L1 adipocytes by incubation with relatively high concentrations of insulin and glucose. Since caffeine has been shown to inhibit adipogenesis in 3T3-L1 cells, we wanted to test the effects of caffeine on untreated mature 3T3-L1 adipocytes, and insulin resistant 3T3-L1 adipocytes. 3T3-L1 adipocytes were grown to maturity following standard methods involving treatment with IBMX, dexamethasone, and insulin. At maturity, insulin resistance in adipocytes will be induced using insulin/glucose feeding, while control adipocytes will be untreated. Insulin resistance in adipocytes will be demonstrated by decreased levels of phosphorylation of Akt. Both control and insulin resistant adipocytes will be treated with 0.5 mM, 1.0 mM, and 5.0 mM caffeine. Western blot analysis will be used to examine caffeine-induced differences in phosphorylation of Akt between control and insulin resistant adipocytes. Caffeine-induced changes in lipid accumulation and cell morphology will be also be examined with microscopy and Oil Red O staining. We believe that caffeine treatment will cause proportionally greater inhibition of Akt phosphorylation and lipid accumulation in insulin resistant adipocytes compared to control adipocytes. The results of this study may serve as a model system for potential caffeine-based treatments of Type 2 diabetes.

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Apr 24th, 4:00 PM Apr 24th, 5:00 PM

The Effects of Caffeine on Insulin Resistant 3T3-L1 Adipocytes

The 3T3-L1 cell line is a commonly used model system in studies on obesity and adipogenesis due to the ease and speed that 3T3-L1 fibroblasts can be differentiated into mature adipocytes. Previous studies have shown that insulin resistance can be induced in mature 3T3-L1 adipocytes by incubation with relatively high concentrations of insulin and glucose. Since caffeine has been shown to inhibit adipogenesis in 3T3-L1 cells, we wanted to test the effects of caffeine on untreated mature 3T3-L1 adipocytes, and insulin resistant 3T3-L1 adipocytes. 3T3-L1 adipocytes were grown to maturity following standard methods involving treatment with IBMX, dexamethasone, and insulin. At maturity, insulin resistance in adipocytes will be induced using insulin/glucose feeding, while control adipocytes will be untreated. Insulin resistance in adipocytes will be demonstrated by decreased levels of phosphorylation of Akt. Both control and insulin resistant adipocytes will be treated with 0.5 mM, 1.0 mM, and 5.0 mM caffeine. Western blot analysis will be used to examine caffeine-induced differences in phosphorylation of Akt between control and insulin resistant adipocytes. Caffeine-induced changes in lipid accumulation and cell morphology will be also be examined with microscopy and Oil Red O staining. We believe that caffeine treatment will cause proportionally greater inhibition of Akt phosphorylation and lipid accumulation in insulin resistant adipocytes compared to control adipocytes. The results of this study may serve as a model system for potential caffeine-based treatments of Type 2 diabetes.