Figure 1

Fabrication of the insulin resistance-induced 3D co-culture model. For the 3D co-culture system, 3D scaffolds were fabricated using 3T3-L1 preadipocytes with or without 1% macrophages. (A) Fluorescent images of 3D mono-cultured 3T3-L1 preadipocytes at 7 days using a live/dead cell assay (magnification: 100×). Live cells stained green and dead cells stained red. (B) The protein concentrations of adipogenesis-related markers, such as CCAAT/enhancer binding protein alpha (C/EBPα), peroxisome proliferator-activated receptor gamma (PPARγ), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), fatty acid binding protein 4 (FABP4), perilipin (PLIN), glucose transporter type 4 (GLUT4) and beta actin (β-actin), were assessed by Western blotting in the 2D and (C) 3D mono-/co-cultured model. (D) Fluorescent image of 3D mono-/co-cultured adipocytes by BODIPY lipid staining (magnification: 100×). (E) Level of glucose uptake in the 3D culture system. Results are expressed as the means ± S.E.M. (n = 4/group) for duplicate experiments. ^###p < 0.001 vs insulin-treated 3D preadipocytes, ***p < 0.001 vs insulin-treated 3D adipocytes. Statistical significance was determined using the Student’s t-test or a one-way analysis of variance followed by Tukey’s multiple-comparison test; p < 0.05 was considered to be statistcally significant. Full-length blots are presented in Figs S1 and S2 in the Supporting Information.