Patient PBMC differentiation to macrophages

After cryorecovery, plate PBMCs at a final concentration of 3 x 10⁶/mL in a poly-D-lysine coated, 96-well plate, resulting in 3x10⁵ cells per well.

Incubate PBMCs at 37°C for 2-hours to allow adherent cells to adhere.

Wash cells washed 2 x sterile PBS to wash off non-adherent cells, and incubate in growth media

containing 20ng/mL of human macrophage-colony stimulating factor (hM-CSF) for 5 days, with a full media change on day 3 to replace nutrients.