Project description:We used microRNA expression analysis to identify which microRNAs are expressed in the stromal vascular fraction (SVF) of visceral white adipose tissue (visWAT) to identify of microRNA expression patterns are changed upon CL-316,243 treatment Agilent mouse miRNA microarray (Cat. No. G4872A-046065, Agilent Technologies) was performed on the SVF of visWAT of 8-week-old C57/BL6 mice treated with CL 316243 or Vehicle (PBS) for three days. Isolated miRNA and total RNA was used for array analysis by Shanghai Biotechnology Corporation. Normalization and analysis for differentially expressed genes were performed using robust multi-array analysis and significance analysis of microarrays (SAM) via R statistical software packages “oligo” and “samr”. Several up- and downregulated microRNAs could be identified and were further varified with qPCR and compared with mRNA expression levels of their main targets.
Project description:A single cell suspension was generated from B3-adrenergic agonist (CL; 1 mg/kg/day) for 4 days and saline mouse SVF, and single-cell mRNAseq libraries were generated with Drop-Seq. A single cell suspension was generated from mouse mature adipocyte nuclei under the following conditions: RT, 24hrs @ 4 °C , 48hrs @ 4 °C, 4days @ 4 °C and B3-adrenergic agonist (CL; 1 mg/kg/day) and single-cell mRNAseq libraries were generated with 10X genomics.
Project description:Single cell sequencing of stromal vascular fraction (SVF) under B3-adrenergic agonist stimulation and mature adipocytes under cold exposure and B3-adrenergic agonist stimulation
Project description:We hypothesize that the combination of mechanical loading with hypoxia culture and TGF-β3 growth factor withdrawal will promote stable, non-hypertrophic chondrogenesis of hBM-MSC embedded in an HA-hydrogel. To this end, we first assessed static hypoxia culture with growth factor withdrawal against static normoxia (20% O2) culture at the global transcriptome and tissue matrix level. We then assess two modalities of mechanical loading (dynamic compression, DC and cyclic hydrostatic pressure, CHP) with growth factor withdrawal against static culture, all under hypoxia. Results showed that Mechanical stimulation and TGF-β3 withdrawal under hypoxia promoted a strong chondrogenic and non-hypertrophic phenotype of hBM-MSC.
Project description:microRNA expression in human monocyte-derived DCs following stimulation with NOD2 ligand MDP, TLR2 ligand Pam3CSK4, or both. 4 condition experiment with two timepoints, and 4 biological replicates. Conditions: unstimulated; MDP stimulated; Pam3CSK4 stimulated; MDP + Pam3CSK4 stimulated. Timepoints: 4 hours and 24 hours.
Project description:We found deletion of Foxp4 in SVF cells would inhibit beige adipocyte differentiation and thermogenesis.In order to investage the binding sites of Foxp4 on genome of SVF cells, we isolated SVF cells from ingunal adipose tissues and did Foxp4 and H3K27ac ChIP-Seqs after two days' browning differentiaion.
Project description:Obesity has become a global health problem. Brown adipose tissue (BAT), specialized for energy expenditure through thermogenesis, potently counteracts obesity. Recently, BAT is also identified in human adults. We found that Lgr4 homozygous mutant (Lgr4m/m) mice display reduced adiposity and exhibit brown-like adipocytes in their WAT depots with higher expression of uncoupling protein 1 (Ucp1). Furthermore, Lgr4 ablation potentiates brown adipocyte differentiation from stromal vascular fraction (SVF) of epididymal WAT (eWAT) in vitro. We used microarrays to emxamine the gene expression profiles of the brown-like adipocytes differentiated from SVF of wild-type and Lgr4 mutant mice. We identified distinct gene expression profiles of these two groups. To demonstrate Lgr4 ablation can potentiate the differentiation of SVF from eWAT toward brown-like adipocytes in vitro, we isolated SVF from epididymal white adipose tissue (eWAT) of wild-type (WT) and Lgr4 mutant mice. We then plated SVF cells in 12-well plate, and differentiated them to brown adipocytes, followed by RNA extraction and hybridization with Affymetrix microarrays.
Project description:We determined the gene signatures of adipocytes differentiated from lipoma SVF. Furthermore, single-cell RNA-sequencing was performed for SVF from inherited (familial) and sporadic lipoma.