Gene expression profile of the capillaries with the surrounding cells from the ossification front of the juvenile and adult mouse femurs. Gene expression profile of the capillaries with the surrounding cells from the ossification front of the juvenile and adult mouse femurs

We performed RNA-Seq and compared expression levels of genes of endothelialal cells together with the surrounding cells derived by laser microdissection (LMD) from 4- and 12-week old ossification fronts of Tg(TIE2GFP)287Sato/J (endothelial cell reporter) mice. The aim of the expriments was to dissect the age-related changes in vascular and perivascular microenvironments during active bone growth and maturation. Overall design: We developed a protocol to cut single capillaries with perivascular cells from undecalcified and unfixed bones of endothelial cell-specific GFP-reporter mice with a high RNA preservation rate for futher NGS. This protocol allowed us to analyse purely unmanipulated snap-frozen trancriptome of cells of interest. We performed laser microdissection (LMD) of single capillary with associated surrounding cells in the ossification front (OF) of juvenile, actively growing (4-week-old) and adult, mature (12-week-old) mice, followed by transcriptome comparison.