Anti human cd3 functional grade purified

The PBMCs (Allcells, Cat: PB005F) were resuspended in CTS^TM AIM V^TM SFM medium (GIBCO, Cat: A3021002) at a density of 1 × 10⁶ cells/ml, while 50 ng/ml anti-human CD3 functional grade purified (eBioscience, Cat: 160037–85) and 50 ng/ml anti-human CD28 functional grade purified (eBioscience, Cat: 160288–85) were added to activate T lymphocytes. The cells were then incubated at 37°C with 5% CO2 for 48 h. Concentrated lentiviruses were added to the cultured cells at a multiplicity of infection of 5, while 200 unit/ml of IL-2 (Shandong Quangang Pharmaceutical, Cat: S20020004) and 4 µg/ml polybrene (Sigma, Cat: h9268-5 g) were added and mixed well. The mixture was then incubated at 37°C with 5% CO₂ for 6–8 h, followed by centrifugation at 300 g for 5 min and replacement of the medium with IL-2-containing fresh CTS^TM AIM V^TM SFM medium. Fresh CTS^TM AIM V^TM SFM medium (with IL-2) was added every 2–3 days. Cell density was maintained at approximately 1 × 10⁶ cells/ml and amplified for 14 days.