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.