Project description:Identification of differentially expressed genes in young (3 month old) versus aged (24 month old) mouse bone marrow derived endothelial cells. Comparison of genes differentially expressed in bone marrow derived endothelial cells of young mice with conditional deletion of mTOR within vascular endothelium.
Project description:Aged hematopoietic stem cells (HSCs) display myeloid-biased differentiation and reduced regenerative potential. In this study, we uncover that P-selectin (Selp) marks a subset of aged HSCs with reduced repopulation capacity. This population of HSCs expresses a prominent aging transcriptome. Overexpression of Selp in young HSCs impaired long-term reconstitution potential and repressed erythropoiesis. We show that IL-1β is elevated in aged bone marrow and administration of IL-1β induces expression of Selp and other aging-associated genes in HSCs. Finally, we demonstrate that transplantation of aged HSCs into young recipients restores a young-like transcriptome, specifically by repressing pro-inflammatory pathways, highlighting the important role of the bone marrow microenvironment in HSC aging.
Project description:To describe the protein profile in hippocampus, colon and ileum tissue’ changing after the old faeces transplants, we adopted a quantitative label free proteomics approach.
Project description:ATAC-seq profiling of Nfat5 KO and wild type macrophages derived from bone marrow (primary cells), treated or not with Lipopolysaccharide (LPS).
Project description:We performed joint single-nucleus RNA (snRNA-seq) and ATAC (snATAC-seq) sequencing on hematopoietic stem and progenitor cells (LSK) isolated from mouse bone marrow of mice of different ages and dietary treatments. Mice were either fed ad libitum continually until bone marrow isolation at young (yAL) or old (oAL) age, or underwent a mild dietary restriction (DR; 30% reduction in food intake by weight) for the two weeks prior to bone marrow isolation at young (yDR) or old (oDR) age to investigate early response to nutrient deprivation. Each of these four samples contained pooled cells from two mice.
Project description:Total RNA of B-MSCs from young and old mice were compared using Agilent microarrays. Bone marrow cells from old and young mice were labeled and sorted.
Project description:Purpose: The goal of this study is to investigate the gene expression in aged (18 month old) and young (3 month old) bone marrow-derived monocytes. Methods: Bone marrow was harvested by flushing the femurs and tibias of young (3 month old) and aged (18 month old) mice in PBS with 2% fetal bovine serum (FBS) on ice. Cells were isolated from bone marrow by density gradient centrifugation using Lympholyte (Cedarlane, CL0531). After centrifugation at 1300 g for 20 min, bone marrow cells were collected from the interface and washed in Ca2+/Mg2+ - free HBSS. The cells were dissociated to a single-cell suspension by filtering through a 70-µm nylon mesh. For monocyte isolation, a suspension of bone marrow cells was stained with Alexa 700 anti-CD3 (1:100, BioLegend, 100215), PE-Cy7 anti-CD19 (1:100, BioLegend, 115520), FITC anti-CD11b (1:100, BD Pharmingen, 553310) and APC anti-Ly6c antibodies (1:100, BioLegend, 128015) in PBS with 0.5% FBS for 30 min. CD11b+Ly6c+CD3-CD19- monocytes were isolated with a BD FACSAria cell sorter. Total RNA was extracted and subjected to bulk RNA-seq using Illumina HiSeq 2000. Results: Expression profiles of the transcriptome are compared between bone marrow-derived monocytes isolated from aged (18 month old) and young (3 month old) mice. Conclusions: Significant differences in the expression levels of genes are noted between the two age groups.
