An Adipocyte Cell Culture Model to Study the Impact of Protein and Micro-RNA Modulation on Adipocyte Function
| Autor: | Jager, Jennifer, Gaudfrin, Mélanie, Gilleron, Jérôme, Cormont, Mireille, Tanti, Jean-François, Tanti, Jean‐François |
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| Přispěvatelé: | Université Côte D'Azur, Inserm, Centre Méditerranéen de Médecine Moleculaire( C3M), Team « Cellular and Molecular Pathophysiology of Obesity and Diabetes », Tanti, Jean-François |
| Rok vydání: | 2021 |
| Předmět: |
General Chemical Engineering
Cell Culture Techniques General Biochemistry Genetics and Molecular Biology chemistry.chemical_compound Mice Insulin resistance Adipocyte 3T3-L1 Cells microRNA medicine Adipocytes Animals Humans [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism cell culture biology General Immunology and Microbiology Chemistry General Neuroscience micro-RNA (miR) RNA Cell Differentiation Transfection small interfering RNA (si-RNA) [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism medicine.disease Cell biology Insulin receptor MicroRNAs transfection Diabetes Mellitus Type 2 Cell culture Lipogenesis biology.protein |
| Zdroj: | Journal of visualized experiments : JoVE Journal of visualized experiments : JoVE, JoVE, 2021, ⟨10.3791/61925⟩ |
| ISSN: | 1940-087X |
| Popis: | Alteration of adipocyte function contributes to the pathogenesis of metabolic diseases including Type 2 diabetes and insulin resistance. This highlights the need to better understand the molecular mechanism involved in adipocyte dysfunction to develop new therapies against obesity-related diseases. Modulating the expression of proteins and micro-RNAs in adipocytes remains highly challenging. This paper describes a protocol to differentiate murine fibroblasts into mature adipocytes and to modulate the expression of proteins and micro-RNAs in mature adipocytes through reverse-transfection using small-interfering RNA (siRNA) and micro-RNA mimicking (miR mimic) oligonucleotides. This reverse-transfection protocol involves the incubation of the transfection reagent and the oligonucleotides to form a complex in the cell culture plate to which the mature adipocytes are added. The adipocytes are then allowed to reattach to the adherent plate surface in the presence of the oligonucleotides/transfection reagent complex. Functional analyses such as the study of insulin signaling, glucose uptake, lipogenesis, and lipolysis can be performed on the transfected 3T3-L1 mature adipocytes to study the impact of protein or micro-RNA manipulation on adipocyte function. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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