Single-Cell Transcriptomics on Young and Aged Bone Marrow Megakaryocyte Progenitor Cells

Aging associates with platelet hyperreactivity and thromboinflammation, which markedly contribute to cardiovascular disease and thrombotic events. Here, we studied the characteristics of megakaryocyte progenitor cells (MkP) in a mouse model of aging. Single-cell RNAseq was performed on bone marrow samples from young (3 months) and old (>24 months) male and female mice. Overall design: Young (3-month-old) and old (>24-month-old) C57BL/6 mice (6 females and 6 males per group) were purchased from Jackson Laboratory. All animal care and experiments were approved by the Institutional Animal Care and Use Committee of the University of Zurich. Femora and Tibiae were harvested and cleaned of muscle and tissue. Bone marrow (BM) was isolated and erythrocytes were removed by incubation with RBC lysis buffer. To enrich MkP cells, BM cells were stained with CD41+ and CD42d+ antibodies and sorted using MACS, followed by a BD FACS Aria III 4L. scRNA-seq dataset for each sample was generated using the Chromium system (10X Genomics). Libraries were sequenced on the Illumina NovaSeq 6000 platform. Raw data files were processed using the Cell Ranger 7.2.0 pipeline.