Atherosclerotic Plaque Cell Dissociation

Atherosclerotic plaque tissues were washed with Dulbecco's phosphate-buffered saline (DPBS, Sigma-Aldrich, United States) within 1 hour after surgery. Each specimen was digested at 37°C for 1 hour using miscible liquids that contained collagenase type IV (GIBCO, Gaithersburg, United States), DNase (Sigma, DN25), hyaluronidase (Sigma, H3506), collagenase type XI (Sigma, C7657), and collagenase type II (Sigma, C6885). The mixture was then filtered through a 70 μm cell strainer with DPBS and washed with red blood cell (RBC) lysis buffer (BD Biosciences, United States). The dissociated cell suspension was washed once with DPBS and resuspended in 1 mL of staining buffer (DPBS containing 5% fetal bovine serum, ScienCell, United States).