2.1.1. Preparation of Peripheral-Blood Mononuclear Cells (PBMCs)
hemolysis
HEMOLYSIS, COLD
Twenty milliliters of peripheral blood were collected from each participant. The blood was used for mononuclear cell isolation as soon as possible using a Lymphoprep™ Tube (Serumwerk, Bernburg, Germany) following the instructions provided. The red blood cells were removed by treating them with an ammonium chloride solution (eBioscience, San Diego, CA, USA) for 10 min at 25 °C. After hemolysis, the cells were washed twice with cold D-PBS (Fujifilm, Tokyo, Japan), counted, and used as PBMCs for the experiments.