Single-cell RNA-seq of vascular endothelial cells in the mouse model of vascular injury
Single-cell RNA-seq of vascular endothelial cells in the mouse model of vascular injury
In the oxygen-induced retinopathy (OIR) model, wild-type C57BL/6J mice were exposed to 75% oxygen (hyperoxia) from P7 to P12 and returned to normal oxygen (normoxia) at P12. Retinal tissues and optic nerves were dissected from OIR and normoxic (control) mice at P12, P17, and P25. Tissues were digested with dispase I, type I collagenase, and type II collagenase. Cells were stained with anti-mouse CD31 and CD45 antibodies. Dead cells were stained with propidium iodide and excluded. CD31+CD45- vascular endothelial cells (ECs) were isolated by fluorescence-activated cell sorting (FACS).  ECs were then loaded into the 10X Chromium Single Cell 3′ Kit (v3) to obtain 5,000 cells per well, and library preparation was performed according to the manufacturer's protocol.  ECs were then loaded into the 10X Chromium Single Cell 3′ Kit (v3) to obtain 5,000 cells per well, and library preparation was performed according to the manufacturer's protocol.