Easysep direct human pbmc isolation kit

Whole blood was collected into 10-mL EDTA tubes from 8 healthy subjects. Written informed consent was received from donors prior to the study. This study was approved by the ethics committee of Shenzhen Zhongshan Obstetrics & Gynecology Hospital (formerly Shenzhen Zhongshan Urology Hospital) and followed the tenants of the Declaration of Helsinki. Human PBMCs (peripheral blood mononuclear cells) were purified with the EasySep™ Direct Human PBMC Isolation Kit (STEMCELL Technologies). MSC-PBMC coculture was performed as described previously with modifications [8 (link), 31 (link)]. Briefly, PBMCs were stimulated with Dynabeads^® Human T-Activator CD3/CD28 (Thermo Fisher Scientific) for 24 h and then cocultured with purified CD317⁺ or CD317⁻ MSCs (20 × 10⁴ PBMCs vs. 5 × 10⁴ MSCs) for 72 h. Cell proliferation was assessed with the Cell Proliferation Kit I (Roche) and quantified by an automated microplate reader (Bio-Rad) at 570 nm.

Written informed consent was obtained from patients involved in the study under the supervision of the local Ethical Committee of Shanghai Tenth People’s Hospital. Throughout the study, the guidelines of the Declaration of Helsinki were observed. 10 healthy volunteers and 29 patients who survived ST-segment elevation MI participated in our study from January 2023 to April 2023. Blood samples from MI patients were collected at day 1 and day 5 respectively. MI was defined according to the Guideline for the Management of ST-Elevation Myocardial Infarction [40 (link)]. Detailed medical and family history of the healthy volunteers was obtained, while none had documented cardiovascular disease, severe kidney or liver disease, or malignancy. All participants’ blood samples were collected after an overnight fast following hospital admission. Basic characteristics were shown in Supplementary Table 1. The PBMCs were extracted using the EasySep™ Direct Human PBMC Isolation Kit (19654, STEMCELL Technologies) according to the manufacturer’s instructions. Information on TREM2’s primer was presented in the Supplementary Table.