Apc anti human cd45ra

We extracted peripheral blood mononuclear cells (PBMCs) by utilizing lymphoprep gradient density centrifugation (AS1114544, Axis-Shield, Norway). PE anti-human CD45RO (304206, Biolegend, USA), APC anti-human CD45RA (304112, Biolegend), FITC anti-human CD8 (100706, Biolegend), PE-cy5 anti-human CD4 (CD45-T2-100, Alpha Diagnostic International), Alexa Fluor^® 488 anti-human CD25(356115, Biolegend) and PerCP anti-human CD3(344813, Biolegend) were used to stain PBMCs, which are co-cultured with OSRC-2 for 48 hours. CD45RO+CD8+ T lymphocytes and CD45RA+CD8+ T lymphocytes were extracted by BD FACSAria II (Becton Dickinson, USA). The fineness of all extracted PBMCs conventionally surpassed 90%. We assembled and suspended 1×10⁶ cells in phosphate-buffered saline (PBS) and then cultured for 5 min at 4°C. Subsequently, the isolated cells were cultured on crushed ice for 0.5 hour in 50 μl of staining solution with 1 μg/ml of corresponding conjugated antibodies. The stained cells were rinsed twice with the pre-cooling PBS. All the samples were examined utilizing the FACS Calibur flow cytometer (BD) and data were analyzed utilizing CellQuest software (BD).