Simple Analysis of Lipid Inhibition Activity on an Adipocyte Micro-Cell Pattern Chip

Abstract

Polydimethyl-siloxane (PDMS) is often applied to fabricate cell chips. In this study, we fabricated an adipocyte microcell pattern chips using PDMS to analyze the inhibition activity of lipid droplets in mouse embryo fibroblast cells (3T3-L1) with anti-obesity agents. To form the PDMS based micropattern, we applied the micro-contact printing technique using PDMS micro-stamps that had been fabricated by conventional soft lithography. This PDMS micro-pattern enabled the selective growth of 3T3-L1 cells onto the specific region by preventing cell adhesion on the PDMS region. It then allowed growth of the 3T3-L1 cells in the chip for 10 days and confirmed that lipid droplets were formed in the 3T3-L1 cells. After treatment of orlistat and quercetin were treated in an adipocyte micro-cell pattern chip with 3T3-L1 cells for six days, we found that orlistat and quercetin exhibited fat inhibition capacities of 19.3% and 24.4% from 0.2 μM of lipid droplets in 3T3-L1 cells. In addition, we conducted a direct quantitative analysis of 3T3-L1 cell differentiation using Oil Red O staining. In conclusion, PDMS-based adipocyte micro-cell pattern chips may contribute to the development of novel bioactive compounds.

Caffeic and Chlorogenic Acids Synergistically Activate Browning Program in Human Adipocytes: Implications of AMPK- and PPAR-Mediated Pathways

“…Initial evaluation of the effect of CA and CGA on adipogenesis was performed via the estimation of intracellular lipids with an Oil red O assay. Despite the fact that the orlistat (ORST) mechanism of action is through gastric/pancreatic lipase inhibition and its direct effect on adipocytes is equivocal [ 50 ], we have employed it as reference drug as it is approved and widely prescribed in anti-obesity therapeutic regimens [ 51 ]. A dose-dependent decrease in lipid accumulation was observed in the cells treated with both phenolic acids ( Figure 2 A), which was confirmed by the spectrophotometric measurements of the extracted dye ( Figure 2 B).…”

“…Transdifferentiation of white adipocytes towards the brown-like phenotype could be induced via several mechanisms, most of which appeared to be PGC-1α/PPAR-dependent. PGC-1α stimulation induces mitochondrial biogenesis and, as a result, increases UCP1 expression in the mitochondria [ 17 , 20 , 49 , 50 , 51 ]. Additionally, browning promotion in WAT is characterized by upregulation of a specific beige marker such as that of T-box 1, tumor necrosis factor receptor superfamily member 9 (CD137), cell death activator CIDE-A (CIDEA) and pyruvate dehydrogenase kinase isoform 4 (PDK4) [ 17 , 19 , 28 ].…”

Molecular Characterization of a Novel Cold-Active Hormone-Sensitive Lipase (HaHSL) from Halocynthiibacter Arcticus

“…In summary, this work provides a molecular understanding of a novel cold-active Ha HSL for potential use in a large variety of biotechnological applications. Based on its unique property, such as high stability in detergents and a cold-active property, Ha HSL could be used as a molecular platform in protein engineering for the preparation of valuable biochemicals or drug intermediates [33,34]. The determination of the Ha HSL structure will allow for a direct comparison to other HSL family members, thus providing insights on the mechanism of action of Ha HSL.…”

Biofabrication of vascularized adipose tissues and their biomedical applications

“…163 With these models, the dose dependent effect of lipid droplet inhibition activity in the presence of anti-obesity agents, for instance Orlistat and Quercetin, can be assessed in 3T3-L1 cells. 165 Also, Rogal et al presented a system that integrates human primary mature adipocytes into a perfused microfluidic chip with their WAT on chip system, where basic and physiological cellular responses as well as drug responses can be monitored. 163 Yang et al designed 3D adipose microtissues by differentiating human-derived ADSCs into mature adipocytes in a microfluidic system.…”