RNAseq for subpopulations of adipose tissue stromal cells from mouse inguinal adipose tissue
Ontology highlight
ABSTRACT: We performed bulk RNA sequencing of several subpopulations of adipose tissue stromal cells to better understand the function of preadipocyte subpopulation (P1 and P2) in mouse inguinal adipose tissue. P1 and P2 cells isolated from inguinal adipose tissue from male and female C57/B6 mice, collected by FACS. P1 cells are CD31-CD45-TER119-SCA1+CD55+VAP1-CD142- cell population, and P2 cells are CD31-CD45-TER119-SCA1+CD55-VAP1+CD142- cell population. 50,000 cells were collected for each populations.
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:To validate that the robustness of Aregs' (CD142+ ASPCs') molecular identity regardless of the antibody used, we performed transcriptomic profiling of freshly isolated 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+ CD142+/− cells of the mouse subcutaneous stromal vascular fraction using FACS.
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:Adipose stem and progenitor cells, defined as live CD45- CD31- Ter119- CD140a+ Sca1+ PDPN+, were sorted from the epididymal adipose tissue of three wild type 12 week old male mice for transcriptional analysis.
Project description:To explore the molecular identity of mouse adipose stem and progenitor cells (ASPCs) and more specifically of CD142+ ASPCs (Aregs), we performed bulk RNA-seq on freshly isolated total, CD142− and CD142+ ASPCs of new-born pups (P0), post-natal day 16 mice (P16), 4 week-old (4wo), 7wo and 11wo mice. ASPCs were collected as Lin− (CD31− CD45− TER119−) CD29+CD34+ SCA1+ cells of the mouse subcutaneous stromal vascular fraction using FACS.
Project description:We have identified two vessel forming mesenchymal stromal cell populations. Population 1 (P1) is CD45-Ter119-Tie2+PDGFRa-CD31+CD105highSca1low and gives rise to stunted vessels (incomplete tubular structures) in a transplant setting, and Population 2 (P2) which is CD45-Ter119-Tie2+PDGFRa+CD31-CD105lowSca1high and forms stunted vessels and fat. To test the homogeneity and signal interactions of these populations we performed 10X single cell RNA-sequencing on them.
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:Population control for the scRNA-seq based analysis a well-established fraction of mouse subcutaneous adipose-derived stromal vascular fraction (SVF) cells that is generally considered to harbour adipogenic stem and progenitor cells (ASPCs). We collected Lin- (CD31- CD45- TER119-) CD29+ CD34+ SCA1+ cells from the mouse subcutaneous SVF of transgenic mice, in which red fluorescent protein (RFP) is induced in Dlk1-expressing cells upon feeding with tamoxifen. While CD29, CD34, and SCA1 are generally expected to enrich for stem cells, DLK1 has previously been suggested to specifically mark pre-adipocytes.
Project description:We analysed a well-established fraction of mouse subcutaneous adipose-derived SVF cells that is generally considered to harbour adipose stem and progenitor cells (ASPCs). To study the molecular characteristics and the subpopulation structure of ASPCs, we performed three replicate scRNA-seq experiments. We collected Lin- (CD31- CD45- TER119-) CD29+ CD34+ SCA1+ cells from the mouse subcutaneous SVF of transgenic mice, in which red fluorescent protein (RFP) is induced in Dlk1-expressing cells upon feeding with tamoxifen. While CD29, CD34, and SCA1 are generally expected to enrich for stem cells, DLK1 has previously been suggested to specifically mark pre-adipocytes. The submission includes the raw data of all sequenced cells, while we only processed 208 high quality single cells. RFP status is indicated in the processed files.
Project description:We have used SmartSeq2 to sequence single phenotypic skeletal stem cell (SSCs) populations purified via FACS from adult mouse long bones. Two putative SSC populations were isolated based on their previously reported surface marker profiles. An osteochondrogenic SSC (ocSSC, CD45-CD31-Tie2-Ter119-CD51+6C3-Thy1-CD105-) and a perivascular SSC (pvSSC, CD45-CD31-Pdgfra+Sca1+CD24+) were investigated.