Platelet Aggregation Assay

Blood samples were obtained with the agreement and approved by the Peking University People's Hospital Medical Ethics Committee (2018PHB210-01) from healthy volunteers without hematological history, platelet or coagulation dysfunction, or taking drugs that may influence hematological function. Platelet-rich plasma (PRP) were obtained from anticoagulated whole blood by centrifugation at 800 rpm for 10 min at room temperature. PRP on the top were transferred to a clean Eppendorf tube. Platelet-poor plasma (PPP) were obtained from re-centrifuging the remaining blood samples at 3,000 rpm for 10 min at room temperature. PRP with the same volume were mixed with the conditioned media derived from sheared HUVECs or saline solution. PPP were mixed with saline solution. Platelet aggregation was triggered by the addition of adenosine diphosphate (ADP, 0.2 μM) (Ya et al., 2019; Chaudhary et al., 2020) and were monitored and recorded using an aggregometer (Chrono-Log, Model Number 700, U.S.A).