Efficient and reliable screening of anti-obesity agents on a micro-cell pattern chip

Mesenchymal stem cells (MSCs) have emerged as one of the promising treatment options for Alzheimer’s disease (AD). Although many studies have investigated on the efficacy of MSCs in AD, how MSCs actually change following exposure to the AD environment has not been studied extensively. In this study, we investigated on the potential of AD patient-cerebrospinal fluid (CSF) samples to be used as a formulation of MSCs and its application in AD therapeutics. When Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) were stored in the CSF of AD patients, the stemness of WJ-MSCs was preserved. Furthermore, several genes were upregulated following storage in AD CSF. This signified the therapeutic potential of CSF formulation for AD therapy. Overall, these findings suggest that CSF from AD patients can be an optimal source for MSC formulation.

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“…Previously described methods were used to quantify the differentiation potential between each samples 39–41 .…”

Section: Methodsmentioning

confidence: 99%

Cerebrospinal fluid from Alzheimer’s disease patients as an optimal formulation for therapeutic application of mesenchymal stem cells in Alzheimer’s disease

Lee

Kwon²,

Kim

et al. 2019

Sci Rep

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“…In addition, to eluting the dye, 100 μL of 100% isopropanol was poured into each adipocyte micro-cell pattern chip for 10 min at room temperature on an orbital shaker, and the Oil Red O was completely dissolved in isopropanol. The absorption was measured at 520 nm using a micro plate reader ( Figure 1 B) [ 21 , 22 ].…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2018

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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.

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“…qualitative analysis, for more than 40 years. [2][3][4] Recently, oil red O staining is also being used for quantitative analysis of adipocyte differentiation [5][6][7][8][9][10][11][12][13][14][15] ( Table 1). To enable quantitative measurements, the dye is commonly eluted from the cells using 2-propanol, and absorbance is photometrically determined at or near the absorbance maximum of oil red O, 518 nm.…”

Section: Introductionmentioning

confidence: 99%

Quantitative assessment of adipocyte differentiation in cell culture

et al. 2016

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Adipocyte cell culture is an important tool for mechanistic studies of energy metabolism. Many factors affect the differentiation of adipocytes in culture. Oil red O staining can be used to assess the degree of differentiation. However, the validity of this method for quantitative analysis has not yet been established. Here we show that a protocol with arbitrarily chosen parameters does not measure in the linear range and is not suitable for quantitative analysis (R 2 D 0.077, p D 0.382), and develop and validate an optimized protocol for quantitative oil red O staining of cultured adipocytes. 3T3-L1 preadipocytes and adipocytes are fixed with 4% formaldehyde and stained with 0.2% oil red O solution in 40% 2-propanol for 30 minutes. Dye is eluted with 2-propanol, and absorption of the eluate is measured photometrically at 510 nm. This optimized protocol achieves excellent correlation between defined amounts of differentiated adipocytes on constant-size culture plates and photometric absorption (R 2 D 0.972, p D 6.585E-14). The performance of the method is independent of the culture plates used. Thus, the optimized oil red O staining protocol can be universally employed to quantitatively assess adipocyte differentiation.

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Efficient and reliable screening of anti-obesity agents on a micro-cell pattern chip

Cited by 4 publications

References 38 publications

Cerebrospinal fluid from Alzheimer’s disease patients as an optimal formulation for therapeutic application of mesenchymal stem cells in Alzheimer’s disease

Cerebrospinal fluid from Alzheimer’s disease patients as an optimal formulation for therapeutic application of mesenchymal stem cells in Alzheimer’s disease

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

Quantitative assessment of adipocyte differentiation in cell culture

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