Effects of co-incubation of LPS-stimulated RAW 264.7 macrophages on leptin production by 3T3-L1 adipocytes: a method for co-incubating distinct adipose tissue cell lines

Abstract

Adipose tissue is a major endocrine organ capable of releasing inflammatory adipokines that are linked to changes occurring in the overfed state, where tissue remodeling results in hypertrophic adipocytes that recruit monocytes to infiltrate the tissue and take on an inflammatory phenotype. Increases in macrophage-specific inflammatory mediator levels contribute to the inflamed state and worsen the inflammatory loop between the macrophages and adipocytes. Although most inflammatory adipokines are released by macrophages, adipocytes can also release immunomodulatory adipokines, such as leptin. The objective of this research was to determine if co-incubation of activated macrophages with mature adipocytes, using transwell inserts, affected adipocyte leptin release. We also examined if there were differences in levels of cell-secreted products quantified in cell-conditioned media collected from macrophage-containing (transwell insert) and adipocyte-containing (well) compartments.