Project description:To clarify the impact of redox homeostasis regulation on chromatin accessibility in adipogenic stem and precursor cells during adipogenic differentiation, we performed ATAC-seq on FIL-ASPCs treated with or without N-acetylcysteine.
Project description:To investigate the function of N6-methyladenosine methylome (m6A) in adipose stem and progenitor cells isolated from facial infiltrating lipomatosis (FIL-ASPCs), we analyzed m6A enrichment level in FIL-ASPCs with or without FTO inhibitor (FTO-IN-1) treatment through methylated RNA immunoprecipitation (MeRIP) sequencing.
Project description:To characterize CD142+ ASPCs (Aregs) after exposure to an adipogenic cocktail we performed bulk RNA-seq (using BRB-seq) of total, CD142− and CD142+ mouse adipose stem and progenitor cells (ASPCs), sorted using four different anti-CD142 antibodies. ASPCs were collected as Lin− (CD31− CD45− TER119−) CD29+ CD34+ SCA1+ cells of the mouse subcutaneous stromal vascular fraction using FACS.
Project description:Human adipose stem and progenitor cells (ASPCs) develop into heterogenous cultures of adipogenic and Structural Wnt-regulated Adipose Tissue resident (SWAT) cells upon induction of adipogenic differentiation. In vitro proliferating ASPC and differentiating adipocytes were collected from multiple timepoints to identify the trajectory of cells. Cells from two white depots (subcutaneous abdominal & visceral abdominal) and two brown depots (supraclavicular & perirenal) were used for the study. Progenitors from all 4 depots show similar differentiation trajectories during early differentiation.
Project description:Human adipose stem and progenitor cells (ASPCs) develop into heterogenous cultures of adipogenic and Structural Wnt-regulated Adipose Tissue resident (SWAT) cells upon induction of adipogenic differentiation. To investiagte the function of SWAT cells and compare them with proliferating ASPCs, we enriched the SWAT cells from the adipogenic cells using density gradient centriguation. The enriched SWAT cells were cultured for 24 hours in proliferation media or differentiation media for 24 hours. 4 density gradient centrifugations were performed, resulting in 4 technical replicates per media type. We included proliferating ASPCS (undifferentiated) as control. We then performed gene expression profiling analysis using data from RNA-Seq of 4 samples per condition (proliferating ASPCs, SWAT cells in proliferation media, SWAT cells in differentiation media)
Project description:To study the involvement of key Areg (CD142+ ASPC)-specifics factors in the inhibitory capacity of CD142+ ASPCs on adipogenesis, we performed transcriptomic profiling of CD142− ASPCs exposed to the secretome of CD142+ ASPCs carrying knockdowns of the indicated genes. CD142− ASPCs were co-cultured and co-differentiated (within the transwell set-up) with CD142+ ASPCs in which siRNA-based knockdowns of specific genes were performed. ASPCs were collected as Lin− (CD31− CD45− TER119−) CD29+ CD34+ SCA1+ cells of the mouse subcutaneous stromal vascular fraction using FACS.
Project description:To characterize the newly discovered Areg cells, we sampled at least four replicates of mouse adipose stem and progenitor cells (ASPCs), CD142+ ASPCs and CD142- ASPCs at four distinct time-points: (1) immediately post-sort (D0), (2) after attachment (5 hours after plating, 5H), (3) after culturing (24 hours after plating, D1) and (4) after adipogenic differentiation (day 12 after plating, day 8 after induction of differentiation, D12), and investigated their transcriptome through a highly multiplexed 3 polyA-selective mRNA-sequencing assay.
Project description:Facial infiltrating lipomatosis (FIL) is a congenital disorder characterized by unilateral facial enlargement. Although next-generation sequencing has revealed that the pathogenesis of FIL is associated with phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA) mutations, the underlying molecular mechanisms remain undetermined. We found that the adipose tissue in FIL patients demonstrated tissue infiltration accompanied by adipocytes hypertrophy and increased lipid accumulation. All FIL derived fibro-adipose progenitor cells (FIL-FAPs) harboured PIK3CA mutations. Compared to FAPs obtained from normal subcutaneous adipose tissue, FIL-FAPs exhibited a greater capacity for adipogenesis. Suppression of PIK3CA resulted in a reduction in the adipogenic potential of FIL-FAPs. Through single cell sequencing, we find that FIL-FAPs had higher expression of FKBP5. Inhibiting FKBP5 can impair the adipogenic capacity of FIL-FAPs. We also verified that PI3K-AKT pathway regulating FKBP5 expression. To find the downstream target of FKBP5, we performed RNA-seq of FIL-FAPs treated with FKBP5 inhibitor SAFit2 in DMSO or DMSO alone.
Project description:To characterize murine ASPCs we performed bulk RNA-seq (BRB-seq, Alpern et al., 2019) of total, CD142− and CD142+ (Aregs) mouse adipose stem and progenitor cells (ASPCs). ASPCs were collected as Lin− (CD31− CD45− TER119−) CD29+ CD34+ SCA1+ cells of the mouse subcutaneous stromal vascular fraction using FACS. The cells were sequenced directly after sort or after expansion.
Project description:We demonstrated the multipotent potential and change of cell identity of human adipose tissue-derived progenitors by subjecting cells to adipogenic, chondrogenic, and osteogenic induction. mRNA transcriptomic profiling results demonstrated the detection of lineage-specific markers.