Human t activator cd3 28

A total of 20 mL of peripheral blood, with approximately 5 × 10⁶ PBMCs, was isolated from each participant and subjected to density gradient centrifugation. The PBMCs were separated into five groups (two control groups and three experimental groups), and wear debris was added at a concentration of 20 particles/PBMC [17 (link)]. The negative control was unchallenged, being resuspended in RPMI 1640 medium (Thermo Fisher Scientific), and the positive control group was treated with human T-activator CD3/28 (Thermo Fisher Scientific), which is known to activate human T-cells. The PBMCs were incubated at 37°C and 5% CO₂ for 72 h, and then analyzed by flow cytometry as described below.

Sixteen milliliters of peripheral blood, with an average of 4 × 10⁶ PBMCs, was isolated from each participant and subjected to density gradient centrifugation. The PBMCs were separated into four groups (two control groups and two experimental groups), and wear debris (20 particles per PBMC) was added to the experimental groups, as described in previous studies.^{6,12 (link)} Each type of particle was directly added into the RPMI 1640 medium (Thermo Scientific, Waltham, MA, USA), except for HXLPE, which was premixed with 100% FBS to avoid particle floatation.^{13 (link)} The negative control was unchallenged in RPMI 1640 medium. The positive control group was treated with human T-activator CD3/28 (Thermo Scientific). The PBMCs were incubated at 37 °C with 5% CO₂ for 72 h and then analyzed by flow cytometry.