Three healthy adult blood was obtained from Acibadem Labcell Cellular Therapy Laboratory.
Following the isolation of peripheral mononuclear cells (PBMC) by overlaying blood on Ficollpaque plus (GE Healthcare), the serially diluted (300 U/mL, 100 U/mL, 30 U/mL, 10 U/mL, 3 U/mL) Dornase alfa (Pulmozyme®, Roche, USA) and 420 g/mL MgSO 4 (Magnesium Sulphate 15% Onfarma, TR) were incubated with the PBMCs for 48hr in T cell medium (6% Human AB Serum and 1% Pen/Strep, Texmacs medium). Immune cell subtypes and their activation levels were determined by Miltenyi MACSQuant flow cytometry analysis using CD3-PE, CD19-PE.cy7, CD56-FITC, CD4-Viogreen, CD8-Vioblue, CD25-APC and CD107a-PE.cy5.5 (Miltenyi).
Following the isolation of peripheral mononuclear cells (PBMC) by overlaying blood on Ficollpaque plus (GE Healthcare), the serially diluted (300 U/mL, 100 U/mL, 30 U/mL, 10 U/mL, 3 U/mL) Dornase alfa (Pulmozyme®, Roche, USA) and 420 g/mL MgSO 4 (Magnesium Sulphate 15% Onfarma, TR) were incubated with the PBMCs for 48hr in T cell medium (6% Human AB Serum and 1% Pen/Strep, Texmacs medium). Immune cell subtypes and their activation levels were determined by Miltenyi MACSQuant flow cytometry analysis using CD3-PE, CD19-PE.cy7, CD56-FITC, CD4-Viogreen, CD8-Vioblue, CD25-APC and CD107a-PE.cy5.5 (Miltenyi).