Project description:Visceral adipose tissue (VAT) is studied extensively to understand its involvement in development of metabolic disorders. In this experiment, we attempt to profile different cell types in VAT of a healthy young individual.
Project description:The complex relationship between metabolic disease risk and body fat distribution in humans involves cellular characteristics which are specific to each body fat compartment. We applied single-cell RNA sequencing (scRNA-Seq) to identify these depot-specific differences in the stromal vascular fraction of visceral (VAT) and subcutaneous (SAT) adipose tissue of obese individuals. We characterized multiple immune cells, endothelial cells, fibroblasts, adipose and hematopoietic stem cell progenitors. Subpopulations of adipose-resident immune cells were found to be metabolically active and associated with metabolic disease status including a population of potential dysfunctional CD8+ T cells expressing metallothioneins. Adipocyte progenitors were identified at different stages of adipogenesis where a subset was pronounced in type 2 diabetics. Depot-specific analysis revealed a class of adipocyte progenitors unique to VAT possessing features similar to ‘inducible brown preadipocytes’. Our first generation human scRNA-Seq transcriptome atlas across fat depots provides a new resource to dissect functional genomics of metabolic disease.
Project description:In order to select mRNA transcripts strongly enriched in murine white adipocytes versus brown adipocytes or stromal-vascular fraction, gene expression data of the adipocyte and stromal-vascular fractions of the interscapular brown, inguinal subcutaneous as well as visceral epididymal adipose tissue depots of young adult male C57BL/6 mice housed at constant 23°C ambient temperature were obtained. 18 samples: 3 different adipose tissues separated into stromal-vascular fraction and adipocytes, analyzed in biological triplicates.
Project description:In order to select mRNA transcripts strongly enriched in murine white adipocytes versus brown adipocytes or stromal-vascular fraction, gene expression data of the adipocyte and stromal-vascular fractions of the interscapular brown, inguinal subcutaneous as well as visceral epididymal adipose tissue depots of young adult male C57BL/6 mice housed at constant 23°C ambient temperature were obtained.
Project description:Unlike visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT) can play a protective role against the development of insulin resistance and metabolic dysfunction in obesity. These changes are associated with higher induction of pro-inflammatory programs and macrophage infiltration in VAT compared with SAT in obesity. Moreover, our results suggest that subcutaneous adipose tissue macrophages (ATMs) release small extracellular vesicles (sEVs) that can improve insulin sensitivity, opposite to the effect of visceral ATM sEVs in obesity. This functional difference was associated with changes in the proportion of resident ATMs, as well as proinflammatory, recruited ATMs. To study the mechanism by which adipose tissue inflammation and macrophage infiltration are differentially regulated in SAT vs VAT, we performed single cell RNA sequencing (scRNA-seq) analysis in sorted live CD45+ CD11b+ myeloid cells from stromal vascular fraction (SVCs) of SAT and VAT of mice fed normal chow diet, high fat diet, or SVCs of HFD mice after the diet switch from high fat diet to normal chow diet, or treatment with insulin sensitizing, rosiglitazone.
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.
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.
Project description:An experiment was performed in order to compare directly profiles of stromal vascular fraction of adipose tissue, testis and epidydimis, in order to rule out a possible contamination of the stromal vascular fraction of adipose tissue (SVF) by adjacent tissues (i.e. epididymis and testis). This led to the identification of a list of 2358 SVF-specific probes, which was subsequently used for further investigations. 3 dye-swap pairs, comparing 3 distinct samples of total RNA prepared from: (1) stromal vascular fraction of adipose tissue, (2) testis and (3) epidydimis.
Project description:Adipose tissue from 6 non-obese patients was collagenase treated and adipocytes separated from the stromal vascular fraction(SVF). SVF was then FACS sorted for the following fractions CD45-/CD34+/CD31+ (endothelial), CD45-/CD34+/CD31- (progenitor), CD45+/CD14+ (monocyte/macrophage), CD45+/CD14-(Leukocyte). RNA was isolated from adipocyte, SVF, progenitor, macrophage/monocyte and leukocyte fractions and analyzed on the Affymetrix Human Transcriptome 2.0 array. We also sorted SVF from an additional 13 (10 non-obese, 9 obese) patients and sent progenitor RNA for Affymetrix Human Transcriptome 2.0 array analysis.