Age of the bone marrow dictates clonality of smooth muscle-derived cells in the atherosclerotic plaque [bulk RNA-seq]

ABSTRACT: 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.

ORGANISM(S): Mus musculus

PROVIDER: GSE206429 | GEO | 2022/08/01

REPOSITORIES: GEO