2.9. Peripheral Blood Mononuclear Cell Extraction
Fresh blood was used for the extraction of PBMC to ensure high viability of the cell sample. Ficoll–Paque density gradient media (15 mL) (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) was added to a 50 mL conical tube. In a separate 15 mL conical tube, 7–10 mL of fresh whole blood was mixed with Dulbecco’s Phosphate Buffered Saline (DPBS) in a 1:1 ratio and inverted four times to mix. The blood:DPBS mixture was then slowly layered above the Ficoll–Paque density gradient media to prevent mixing. Once layered, the 50 mL conical tube containing fresh blood and Ficoll–Paque density gradient media was spun down at 400× 