Differentiated Mouse Adipocytes in Primary Culture: A Model of Insulin Resistance
Por:
Ruiz-Herrera, Xarubet, Luzardo-Ocampo, Ivan, de la Escalera, Gonzalo Martinez, Clapp, Carmen, Macotela, Yazmin
Publicada:
1 feb 2023
Resumen:
Insulin resistance is a reduced effect of insulin on its target cells,
usually derived from decreased insulin receptor signaling. Insulin
resistance contributes to the development of type 2 diabetes (T2D) and
other obesity-derived diseases of high prevalence worldwide. Therefore,
understanding the mechanisms underlying insulin resistance is of great
relevance. Several models have been used to study insulin resistance
both in vivo and in vitro; primary adipocytes represent an attractive
option to study the mechanisms of insulin resistance and identify
molecules that counteract this condition and the molecular targets of
insulin-sensitizing drugs. Here, we have established an insulin
resistance model using primary adipocytes in culture treated with tumor
necrosis factor-alpha (TNF-alpha).Adipocyte precursor cells (APCs),
isolated from collagenase-digested mouse subcutaneous adipose tissue by
magnetic cell separation technology, are differentiated into primary
adipocytes. Insulin resistance is then induced by treatment with
TNF-alpha, a proinflammatory cytokine that reduces the tyrosine
phosphorylation/ activation of members of the insulin signaling cascade.
Decreased phosphorylation of insulin receptor (IR), insulin receptor
substrate (IRS-1), and protein kinase B (AKT) are quantified by western
blot. This method provides an excellent tool to study the mechanisms
mediating insulin resistance in adipose tissue.
Filiaciones:
Ruiz-Herrera, Xarubet:
Instituto de Neurobiología, Universidad Nacional Autónoma de México (UNAM), Mexico
Luzardo-Ocampo, Ivan:
Instituto de Neurobiología, Universidad Nacional Autónoma de México (UNAM), Mexico
de la Escalera, Gonzalo Martinez:
Instituto de Neurobiología, Universidad Nacional Autónoma de México (UNAM), Mexico
Clapp, Carmen:
Instituto de Neurobiología, Universidad Nacional Autónoma de México (UNAM), Mexico
Macotela, Yazmin:
Instituto de Neurobiología, Universidad Nacional Autónoma de México (UNAM), Mexico
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