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

Gi Kim,Changhyun Roh,Chang-Soo Lee,Sung-Chan Jang,Su-Jin Yeom,Heon-Ho Jeong

doi:10.3390/biom8020037

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.

Highlights

Obesity, which arises from an imbalance of energy intake and expenditure, has a tremendous socioeconomic impact at an epidemic level
SU-8 photo-resists were purchased from Microchem (Newton, MA, USA); the 0.45 μm syringe filter and Sylgard 184 PDMS prepolymer were purchased from Dow Corning (Midland, MI, USA); and the Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin (PS), and 0.25% trypsin-ethylenediaminetetraacetic acid (EDTA) were purchased from Gibco (New York, NY, USA)
The adipocyte micro-cell pattern chip successfully enabled the selective growth of 3T3-L1 cells onto the cell adhesion region

Summary

Introduction

Obesity, which arises from an imbalance of energy intake and expenditure, has a tremendous socioeconomic impact at an epidemic level. The World Health Organization (WHO) reported that two billion people suffer from obesity [1,2]. Significant research on development of a lipid inhibitor for obesity is required. Only orlistat [(S)-((S)-1-((2S,3S)-3-hexyl-4-oxooxetan-2-yl)tridecan-2-yl) 2-formamido-4-methylpentanoate], an anti-obesity agent, which is commonly used as an anti-obesity medication, has been approved by the Food and Drug Administration (FDA) [5,6]. Orlistat has shown efficacy in reducing obesity, there have been adverse side-effects [7,8]. Screening for novel bioactive compounds is important to prevent or reduce the risks of various obesity complications. Quercetin has been reported to activate the inhibition of the oxidation of low-density lipoproteins in vitro, anti-obesity agents, free radical scavengers, protein kinase enzyme inhibitors, and estrogen receptors [10,11,12]. The demand for a biological screening assay platform has been issued to efficiently analyze these kind of drug candidates

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Results

Conclusion