Primary adipocytes as a model for insulin sensitivity

Abstract

Obesity and its comorbidity insulin resistance lead to the development of chronic metabolic diseases, such as impaired fasted blood glucose and type 2 diabetes. Adipose tissue plays an important role in whole-body glucose homeostasis, particularly in obese individuals; therefore, many in vivo models of type 2 diabetes are obese, such as Lepob/ob and Leprdb/db mice or ZDF rats. Primary adipocytes therefore represent an attractive in vitro model to study insulin-mediated glucose uptake to investigate the mechanisms of insulin resistance and explore the potential insulin-sensitizing properties of new antidiabetic drugs. Primary adipocytes are isolated by collagenase digestion of adipose tissue, Glucose transport is evaluated by the measurement of intracellular uptake of a tracer (D-[U14C] glucose). The uptake of [U-14 C] glucose reflects directly glucose transport. In this chapter, we will describe the protocol for the isolation of primary rodent adipocytes and the measurement of basal and insulin-stimulated glucose uptake.