Peripheral Blood Mononuclear Cell (PBMC) Isolation
Freshly collected samples were diluted in dPBS at room temperature.
Blood samples were overlayered onto Lymphoprep (Stemcell Technologies,
Cambridge, UK) and centrifuged at 800 xg for 10 minutes at room
temperature. The mononuclear cell interphase band containing monocytes
and lymphocytes was aspirated and washed in HBSS medium and resuspended
in RPMI 1640. 2X Freezing Media (10% DMSO, 40% FCS, 50% RPMI 1640)
was added to cell suspensions. Samples were transferred into cryovials
and placed at -80°C. After 24 hours, cryovials were transferred into
liquid nitrogen where they were stored until further use.