Project description:To determine how neutrophils are reprogrammed by lung CD140a+ MCs (mesenchymal cells), we employed lung CD140a+ MCs, and ex-vivo cultured with BM(bone marrow)-derived neutrophils. After the treatment, neutrophils were harvested for RNA extraction and the transcriptional profiles were analyzed by RNA sequencing (RNA-seq).
Project description:To better understand how lung neutrophils transcriptionally differ from other tissue neutrophils, we performed single-cell RNA sequencing (scRNA-seq) on BM, PB and lung neutrophils isolated from naïve (BALBc/J) and 4T1 tumor-bearing mice.
Project description:To investigate the heterogeneity of lung CD140a+ mesenchymal cells (fibroblasts) and identify the specific lung fibroblast subset, we purified CD140a+ lung cells using CD140a-EGFP reporter mice for a higher-resolution scRNA-seq.
Project description:Human fetal dissociates from 19-22 week gestational age were magnetically sorted for CD140a antigen. CD140a-defined OPCs were plated into serum free conditions and allowed to differentiate in the absence of growth factors or mitogens. RNA was extracted from cells immediately following isolation and every day for 4 days. To block differentiation, matched cells were cultured in the presence of PDGF-AA (10ng/ml). This treatment prevents the acquisition of O4-positive oligodendrocyte cell fate and delays MBP mRNA expression by human CD140a-sorted OPCs.
Project description:This array analysis is to study the regulation of target messages’ expression in in vitro cultured murine neutrophils versus miR-223 null neutrophils. Culture media was SILAC-IMDM for MS analysis.
Project description:Glial progenitor cells (GPCs) pervade the human brain. These cells express gangliosides recognized by MAb A2B5, and some but not all can generate oligodendrocytes. Since some A2B5+ GPCs express PDGFa receptor (PDGFRa), which is critical to oligodendrocyte development, we asked if PDGFRa-directed sorting might isolate oligodendrocyte-competent progenitors. We used FACS to sort PDGFRa+ cells from the second trimester fetal human forebrain, based on expression of the PDGFRa epitope CD140a. CD140a+ cells could be maintained as mitotic progenitors that could be instructed to either oligodendrocyte or astrocyte phenotype. Transplanted CD140a+ cells robustly myelinated the hypomyelinated shiverer mouse brain. Microarray confirmed that CD140a+ cells differentially expressed PDGFRA, NG2, OLIG1/2, NKX2.2 and SOX2. Some expressed CD9, thereby defining a CD140a+/CD9+ fraction of oligodendrocyte-biased progenitors. CD140a+ cells differentially expressed genes of the PTN-PTPRZ1, wnt, notch and BMP pathways, suggesting the interaction of self-renewal and fate-restricting pathways in these cells, while identifying targets for their mobilization and instruction.
Project description:We employed GeneChip analysis to investigate the global gene expression profiles of neutrophils from BM Gr-1high/CD48– neutrophils were sorted from the BM of C57BL/6 mice by FACS in two independent experiments
Project description:To gain a better understanding of the role of Interleukin-1β (IL-1β) in lung CD140a+ mesenchymal cells (fibroblasts) modulation, we performed RNA-seq to compare the transcriptomes of IL-1β-treated and control lung CD140a+ mesenchymal cells (fibroblasts).