Sepmate collection tubes

Within 2 h of whole blood collection, PBMCs were isolated from peripheral blood. T cells were enriched using RosetteSep™ Human T‐Cell Enrichment Cocktail (StemCell Technologies, Cat #15061) according to the manufacturer's protocol. Both PBMCs and T cells were collected via SepMate™ collection tubes (StemCell Technologies, Cat #85460), using Lymphoprep™ (StemCell Technologies, Cat #7861) density gradient, and washed twice. All isolated cells were frozen in CryoStor® (StemCell Technologies, Cat #7930) at a controlled rate using Mr. Frosty containers.

Blood was collected in vacutainer® SST tubes (BD), serum was obtained and stored at −20°C for further experiments. PBMC were isolated from blood and collected in vacutainer tubes containing lithium heparin as anticoagulant by density gradient centrifugation with Lymphoprep™ (Stemcell Technologies) in 50 mL SepMate™ collection tubes (Stemcell Technologies) according to manufacturer’s instructions. Briefly, blood was diluted in phosphate buffered saline (PBS), loaded onto Lymphoprep™ and PBMCs were separated by centrifugation at 2000 g for 15 minutes. PBMCs were washed 3 times in PBS, counted and frozen in 90% fetal bovine serum (FBS) with 10% DMSO (Honeywell) in liquid nitrogen.