An Adipocyte Cell Culture Model to Study the Impact of Protein and Micro-RNA Modulation on Adipocyte Function

Description

Alteration of adipocyte function plays a key role in the pathogenesis of metabolic diseases such as insulin resistance and Type 2 Diabetes. This highlights the need to better understand the molecular mechanism involved in adipocyte dysfunction in order to develop new therapies against obesity-related diseases. Manipulating the expression of proteins and micro-RNAs in adipocytes remains highly challenging. This manuscript describes a protocol to differentiate murine fibroblasts into mature adipocytes and, to manipulate the expression of proteins and micro-RNAs in mature adipocytes through reverse transfection using small interfering RNA and micro-RNA mimic oligonucleotides. Functional experiments such as insulin signaling, glucose uptake, lipogenesis, and lipolysis can be performed on the transfected 3T3-L1 mature adipocytes to study the impact of proteins or micro-RNAs manipulation on adipocyte function.

Abstract

International audience

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