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Genome wide analysis of Visceral adipose tissue CD14+ cells from Obese and obese diabetic subjects

ABSTRACT: Identification of the inflammatory signature in visceral adipose tissue CD14+ cells (adipose tissue macrophage) Total RNA obtained from CD14+ cells (Immunoselcted cells from stromal adipose tissue cells) Adipocytes and cells of the stroma vascular fraction (SVF) were obtained by collagenase digestion of adipose tissue. SVF cells were resuspended in endotoxin-free PBS supplemented with 2% FCS and 1 mM EDTA. Isolation of CD14+ using positive selection magnetic beads (Stemcell technologies) and CD4+ cells was performed using positive selection magnetic beads (Stemcell Technologies, Vancouver, Canada). An Illumina (San Diego, CA) RNA amplification kit (NuGEN, BiotinIL Module) was used according to the manufacturer's instructions to obtain biotinlabeled cDNA from 50 ng of total RNA extracted from adipose tissue CD14+ cells.

ORGANISM(S): Homo sapiens

SUBMITTER: Michele Guerre-Millo

PROVIDER: E-GEOD-54350 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Genome wide analysis of Visceral adipose tissue CD14+ cells from obese subjects

Project description:Identification of the inflammatory signature in visceral adipose tissue CD14+ cells (adipose tissue macrophage) Total RNA obtained from CD14+ cells (Immunoselcted cells from stromal adipose tissue cells) Adipocytes and cells of the stroma vascular fraction (SVF) were obtained by collagenase digestion of adipose tissue. SVF cells were resuspended in endotoxin-free PBS supplemented with 2% FCS and 1 mM EDTA. Isolation of CD14+ using positive selection magnetic beads (Stemcell technologies) and CD4+ cells was performed using positive selection magnetic beads (Stemcell Technologies, Vancouver, Canada). An Illumina (San Diego, CA) RNA amplification kit (NuGEN, BiotinIL Module) was used according to the manufacturer's instructions to obtain biotinlabeled cDNA from 50 ng of total RNA extracted from adipose tissue CD14+ cells.

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Genome wide analysis of Visceral adipose tissue CD14+ cells from Obese and obese diabetic subjects

2014-01-24 | GSE54350 | GEO

Genome wide analysis of Visceral adipose tissue CD14+ cells from obese subjects

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Gene expression in canine adipose stromal vascular fraction

Project description:Microarray studies were performed to identify expression patterns by cryopreserved adipose stromal vascular fraction (SVF) preparations that were prepared by three different veterinary stem cell companies from the same source of canine adipose tissue. [Samples 1 - 3] Three equal samples of approximately 20 grams of adipose tissue were shipped on ice to 3 different companies which specialize in adipose-derived veterinary stem cell therapy. All 3 SVF preparations were cryopreserved at their respective facilities. Approximately one month later, cryopreserved samples were retrieved from all companies and transported frozen to the University of Kentucky for gene profiling and viability analysis. [Samples 4 - 11] Groups of cells (samples 5, 7, 9, 11) were treated by the stem cell company with photo-activated platelet-rich plasma (PRP) according to their proprietary procedures in order to enhance regenerative properties of the adipose stem cells. These PRP-treated cells were incubated in the investigator's lab for 6 hours at 37 degrees Celsius to allow for expression of genes that were induced by PRP treatment.

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