Flow Cytometry Analysis

Confluent meningioma cells were resuspended in ice-cold staining buffer (PBS, 2% FBS) at a final concentration of 1 × 10⁵ cells. Cells were stained for 30 min at RT in the dark with the following: CD45-FITC, HLA-DR-PE, CD14-PerCP5.5, and CD44—APC (Becton Dickinson Biosciences, Pharmingen), washed twice with 2 ml of staining buffer and centrifuged at 1500 rpm for 5 min at 4°C. The relevant single isotype controls were used. Data acquisition was collected on 1 × 10⁴ cells on an Accuri flow cytometer (BD Biosciences) and analysis was performed using Flow Jo software v10.0 (FlowJo LLC).