Magnetic cell sorter macs

Frozen PBMC were thawed, washed and incubated with 20μl of CD25 micro beads II, human (Miteny Biotec, Bergisch Gladbach, Germany) in 80μl MACS buffer (Phosphate-buffered saline (PBS) with 0.5% Bovine serum albumin (BSA) and 2 mM EDTA) for 20 minutes at 4°C. Cells were washed and added to a MS column attached to a Magnetic Cell Sorter (MACS) (Milteny Biotec). CD25^- cells were collected as flow through and the CD25⁺ population was collected by detaching the column from the magnetic cell sorter. Cells were washed with MACS buffer and resuspended in AIM-V medium. The purity of the CD25⁻ and CD25⁺cell populations was >95% (S1 Fig).

Frozen PBMC were thawed, washed and incubated with 20 µl of the CD25 micro beads II, human (Miteny Biotec, Bergisch Gladbach, Germany) in 80 µl MACS buffer (Phosphate-buffered saline (PBS) with 0.5% Bovine serum albumin (BSA) and 2 mM EDTA) for 20 minutes at 4°C. Cells were washed and added to MS column attached to Magnetic Cell Sorter (MACS) (Milteny Biotec) where CD25⁻ cells were collected as flow through and the CD25⁺ population was collected by detaching the column from the magnetic cell sorter. Cells were washed with MACS buffer and resuspended in AIM-V medium. The purity of the CD25⁻ and CD25⁺cell populations was >80% (supplementary figure S2A and S2B).