Abstract
Several studies have shown that the detrimental influence of abdominal obesity on metabolic processes is mediated by the intra-abdominal fat depot. Visceral adipose tissue has been shown to be an independent risk factor for coronary heart disease, hypertension, impaired glucose tolerance and Diabetes Mellitus Type 2 (DM2). Diet-induced obesity in mice, primarily of the C57BL/6J strain, is a commonly used method to study the development of insulin resistance as a model for DM2. The white or visceral adipose tissue (here referred to as VAT), especially the fat around the gonads, is a commonly used organ of study in this model, as it accumulates large numbers of lymphocytes in response to diet-induced obesity. The protocol below describes the isolation of lymphocytes from the stromal vascular fraction (SVF) from VAT.
Materials and Reagents
Equipment
Procedure
Representative data
Figure 9. FACS plot of Leukocytes isolated from VAT. Cells were stained with viable dye [Propidium Iodide (PI)] and CD45 antibodies. Gated is for singlets.
Supplementary figures
Supplementary Figure 1a. Gonadal/visceral adipose tissue in female mice Supplementary Figure 1b. Gonadal/visceral adipose tissue in male mice
Recipes
Acknowledgments
When using this protocol, please refer to Wensveen et al. (2015). This work was supported by the European Foundation for the Study of Diabetes (New Horizons Program), the Unity through Knowledge Fund (15/13 to B. P.), the University of Rijeka (13.06.1.1.03 to B. P.), the EU ESFEuropean Social Fund - ES (HR.3.2.01-0263 to B.P.), the Netherlands Organization for Scientific Research (91614029 to F. M. W.) and the European Commission (PCIG14-GA-2013-630827 to F. M. W.).
References
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