Obesity-induced white adipose tissue (WAT) fibrosis is believed to accelerate WAT dysfunction. However, the cellular origin of WAT fibrosis remains unclear. Here, we show that adipocyte platelet-derived growth factor receptor-α-positive (PDGFRα+) progenitors adopt a fibrogenic phenotype in obese mice prone to visceral WAT fibrosis. More specifically, a subset of PDGFRα+ cells with high CD9 expression (CD9high) originates pro-fibrotic cells whereas their CD9low counterparts, committed to adipogen ...[more]
Project description:Obesity-induced white adipose tissue (WAT) fibrosis is believed to accelerate WAT dysfunction. Two progenitor populations could be distinguished in omental white adipose tissue (oWAT) of morbidly obese individuals based on CD9 expression. In addition, the frequency of CD9high progenitors in oWAT correlates with oWAT fibrosis level, insulin-resistance severity and type 2 diabetes. To further gain insight into the functional differences between the CD9high and CD9low progenitor subsets, we performed transcriptomic profiling of FACS-sorted progenitor populations isolated from oWAT of obese individuals. As CD9low progenitors were markedly decreased in glucose-intolerant or diabetic individuals, we investigated seven women with morbid obesity but without diabetes or glucose intolerance, according to the ADA definition. Overall design: Omental adipose tissue from 7 obese women with mean BMI of 42.9±1.7kg/m2 and mean age of 40.7±3.3 year-old was digested with collagenase. CD9high and CD9low progenitors from the stromal vascular fraction were FACS-sorted with CD45- CD31- CD34+ CD44+ PDGFRa+ CD9high and CD45- CD31- CD34+ CD44+ PDGFRa+ CD9low. Total RNA was amplified and labeled using Ovation Pico WTA System V2 (NuGEN) and Encore BiotinIL Module (NuGEN) kits according to the manufacturer's protocol.
Project description:Obesity-induced white adipose tissue (WAT) fibrosis is believed to accelerate WAT dysfunction. However, the cellular origin of WAT fibrosis remains unclear. We showed that adipocyte platelet-derived growth factor receptor-a-positive (PDGFRa+) progenitors adopt a fibrogenic phenotype in obese C3H/HeOuJ (C3H) mice prone to visceral WAT fibrosis. Two progenitor populations could be distinguished in the epididymal white adipose tissue (EpiWAT) of lean C3H mice, based on CD9 expression. In the fibrotic EpiWAT of obese C3H animals, the PDGFRa+CD9low subset is almost completely lost while the density of PDGFRa+CD9high progenitor markedly increase. To further gain insight into the functional differences between the CD9high and CD9low progenitor subsets, we performed transcriptomic profiling of FACS-sorted progenitor populations isolated from EpiWAT of lean C3H mice. Overall design: EpiWAT from 3-month-old C3H mice was digested with collagenase. CD9high and CD9low progenitors from the stromal vascular fraction were FACS-sorted with CD45- CD31- Gp38+ PDGFRa+ CD9high and CD45- CD31- Gp38+ PDGFRa+ CD9high. Total RNA was amplified and labeled using Ovation Pico WTA System V2 (NuGEN) and Encore BiotinIL Module (NuGEN) kits according to the manufacturer’s protocol.
Project description:White adipose tissue regulates metabolism; the importance of this control is highlighted by the ongoing pandemic of obesity and associated complications such as diabetes, atherosclerosis, and cancer. White adipose tissue maintenance is a dynamic process, very little is known about how pharmacologic stimuli affect such plasticity. Combining in vivo lineage marking and BrdU labeling strategies, we found that rosiglitazone, a member of the thiazolidinedione class of glucose-lowering medicines, markedly increases the evolution of adipose progenitors into adipocytes. Notably, chronic rosiglitazone administration disrupts the adipogenic and self-renewal capacities of the stem cell compartment and alters its molecular characteristics. These data unravel unknown aspects of adipose dynamics and provide a basis to manipulate the adipose lineage for therapeutic ends. The goal of this gene expression array was to identify changes in molecular expression in adipose progenitors isolated from mice that underwent two-month rosiglitazone treatment. Adipose SV GFP+ cells (adipose progenitors) were FACS-isolated from adult AdipoTrak mice that had been treated with or without rosiglitazone (0.0075%) for 2 months. RNAs isolated from these cells were used for microarray. Each cohort contains 3-4 mice, each experimental group (-TZD and +TZD) contains 3 cohorts.
Project description:The aim of the project was to compare global gene expression in adipocytes from obese patients and lean controls. Subcutaneous adipose tissue was collected from severely obese patients undergoing bariatric surgery (average body-mass index (BMI) of 45.5 kg/m2 (n = 12, thereof 4 men) and healthy lean patients undergoing hernia repairs (average BMI of 24.2 kg/m2 (n = 12, thereof 7 men), between 27 and 56 years of age. Adipocytes were isolated by collagenase treatment of adipose tissue, followed by filtering and centrifugation. Floating adipocytes were lysed in Qiazol before RNA purification and microarray analysis.
Project description:The study aims to show that Sox9+ Chd1+ mouse embryonic lung progenitors can be isolated and expanded long-term in 3D culture while maintaining their multipotency. in vitro cultured Sox9+ Chd1+ lung progenitors transcriptionally resemble their in vivo counterparts and show significant difference from adult lung epithelial (Cdh1+ and EpCAM+) and non-epithelial (Cdh1- and EpCAM-) cells
Project description:Tissue-resident mononuclear phagocytes (MNPs) in metabolic organs contribute to the regulation of whole body metabolism. CD301b+ MNPs are a subset of MNPs that are found in most peripheral organs including metabolic organs. In a mouse model in which CD301b+ MNPs can be selectively and transiently depleted, we examined the impact of the depletion on gene expression in the white adipose tissue and the liver. Overall design: Male Mgl2-DTR mice or control wild-type mice were treated with diphtheria toxin for 10 days to depleted CD301b+ MNPs. Total RNA was isolated frrom the inguinal and epididymal white adipose tissue and the liver.
Project description:We performed microarray analysis to evaluate differences in the transcriptome of the adipose tissue before and during pregnancy Biopsies of adipose tissue of the subcutaneous gluteal depots, processed for adipose cell isolation, the remining tissue was snap frozen in liquid nitrogen, adipocytes were isolated by digestion with collagenase
Project description:Lin-Sca-1+ adipocyte progenitors from subcutaneous adipose tissue of wild-type and Cited4-/- female mice were cultured under conditions promoting general adipogenic differentiation or beige/brite adipocyte differentiation (treatment with Rosi). Cells were harvested 48 post-induction of differentiation.