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

LPS 刺激的 RAW 264.7 巨噬细胞共培养对 3T3-L1 脂肪细胞产生瘦素的影响:一种共培养不同脂肪组织细胞系的方法

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作者:Cristina Caldari-Torres, Jordan Beck

Background

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

Conclusion

Our results suggest that co-incubating activated macrophages and adipocytes using transwell inserts can result in distinct microenvironments in the different cellular compartments and that separate sampling of these compartments is required to detect the subtle signaling dynamics that exist between these cells.

Methods

Mature adipocytes were co-incubated with control and lipopolysaccharide-stimulated (0.01 mg/ml) murine macrophages, and nitric oxide, interleukin-6, and leptin levels were quantified in the cell-conditioned media from both compartments.

Results

Activation status of the macrophages did not affect leptin release by the adipocytes. We observed higher amounts of leptin in wells compared to transwells. Nitric oxide and interleukin-6 levels were similar between transwells and wells, suggesting that these adipokines travel through the transwell inserts and are reaching equilibrium between the two compartments.

Supplementary Information

The online version contains supplementary material available at 10.1186/s42269-022-00747-7.

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