Data from: Adipose tissue explant culture using PDMS flow chambers: an alternative to static explant culture
Data files
Dec 12, 2025 version files 5.78 MB
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120hr_Post_Chamber.jpg
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120hr_Post_Dish.jpg
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120hr_Pre.jpg
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216hr_Post_Chamber_CD45_only_zoom.png
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216hr_Post_Chamber.jpg
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216hr_Post_Dish.jpg
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216hr_Pre_Chamber_CD45_only_zoom.jpg
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216hr_Pre.jpg
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72hr_Post_Chamber.jpg
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72hr_Post_Dish.jpg
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72hr_Pre.jpg
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Cohen_et_al_compiled_data.xlsx
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README.md
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Abstract
As obesity continues to rise, it is important we can effectively study adipose tissue. However, adipose tissue is notoriously difficult to culture ex vivo due to fragility, bouyancy, and high nutrient demant. To address these challenges, we developed a novel approach using polydimethylsiloxane (PDMS) flow chambers attached to a micro peristaltic pump for culture of primary adipose tissue explants. Our results demonstrate that adipose tissue remains viable and functional in flow chambers, therefore providing an alternative to static explant culture in a dish.
Dataset DOI: 10.5061/dryad.866t1g22t
Description of the data and file structure
All experiments were performed at UCLA in Los Angeles, CA. Flow chambers for culture of primary adipose tissue explants were designed in house and fabricated using polydimethylsiloxane (PDMS) and 0.4 micron polyethylene terephthalate (PET) membranes. Once produced, chambers were placed in 37˚C tissue culture incubators and attached to a mini peristaltic pump to allow for media perfusion. Tissue explants in flow chambers were assessed for viability using resazurin assays and by measurement of lactate dehydrogenase in media effluent. Explants were then treated with isoproterenol and insulin to test for induction and inhibition of lipolysis, respectively, by colorimetric glycerol assay of media effluent. Finally, explants were collected after incubation in chambers and assessed for maintenance of adipocyte structure using confocal microscopy, and maintenance of adipose gene expression by RT-qPCR. Images and gene expression data were also obtained from static cultured explants in a dish for comparison.
Files and variables
File: Cohen_et_al_compiled_data.xlsx
Description: Microsoft excel file containing compiled data from all quantitative experiments in human and mouse tissues: colorimetric resazurin (alamarblue) viability assay, luminescent LDH assay, colorimetric glycerol release assay, and RT-qPCR
cells with value "null" were not applicable to analysis or not collected at the listed time point
Variables (Tab 1,2)
Effluent = reduced alamarblue chamber effluent
Input = alamarblue media (1:10 dilution in RPMI complete media) before infusion to chamber
% reduction = percent reduced resazurin calculated using equation provided by the manufacturer protocol
Variables (Tab 3)
Iso = 10µM isoproterenol treatment
Ins = 100nM insulin treatment
Time 0: media before entering chamber (blank; subtracted from all values)
Variables (Tab 4)
TX-100 = 0.2% Triton TX-100 detergent in RPMI complete media
Variables (Tab 5,6,7)
M199 = Media 199 with earle's salts and penicillin/streptomycin, serum free
RPMI = RPMI media +10% FBS +1% Glutamax +1% sodium pyruvate + 0.1% gentamicin
Pre: sample before processing for culture
Post: 72 hours in chamber (except tab 6: 120 hours)
Dish: 72 hours in culture dish (except tab 6: 120 hours)
RQ: Relative quantification
CT/CQ: critical threshold/quantification cycle
File: 72hr_Post_Dish.jpg
Description: Human adipose tissue imaged via confocal microscopy after 72 hours in culture plate. Red = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 120hr_Post_Chamber.jpg
Description: Human adipose tissue imaged via confocal microscopy after 120 hours in flow chamber. Purple = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 216hr_Post_Chamber.jpg
Description: Human adipose tissue imaged via confocal microscopy after 216 hours in flow chamber. Purple = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 72hr_Post_Chamber.jpg
Description: Human adipose tissue imaged via confocal microscopy after 72 hours in flow chamber. Red = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 216hr_Pre_Chamber_CD45_only_zoom.jpg
Description: Magnified confocal image of CD45+ immune cells in adipose tissue before flow chamber. Green=CD45, Blue=DAPI (nuclei)
File: 72hr_Pre.jpg
Description: : Human adipose tissue imaged via confocal microscopy before processing for culture. Red = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 216hr_Post_Chamber_CD45_only_zoom.png
Description: Magnified confocal image of CD45+ immune cells in adipose tissue after 216 hours in flow chamber. Green = CD45, Blue = DAPI (nuclei)
File: 120hr_Post_Dish.jpg
Description: Human adipose tissue imaged via confocal microscopy after 120 hours in culture dish. Purple = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 216hr_Post_Dish.jpg
Description: Human adipose tissue imaged via confocal microscopy after 216 hours in culture dish. Purple = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 120hr_Pre.jpg
Description: Human adipose tissue imaged via confocal microscopy before processing for culture. Purple = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
File: 216hr_Pre.jpg
Description: Human adipose tissue imaged via confocal microscopy before processing for culture. Purple = Perilipin 1 (adipocytes), Blue = DAPI (nuclei), Green = CD45 (immune cells)
Human subjects data
Human adipose tissue remnant samples were collected by the UCLA Translational Pathology Core Laboratory after obtaining written informed consent to use samples for research purposes which may result in publication (IRB-11-2504). Distribution of samples to our laboratory was de-identified and included no protected health information. Information provided was as follows: age, sex, diagnosis, tissue location, height, weight, BMI.