Vacuette sodium heparin tubes 9 ml

Peripheral venous blood was collected in three Vacuette® sodium-heparin tubes à 9 mL (Greiner Bio-One; Austria) in the morning (8–10 A.M). All participants had been fasting since 10 P.M the night before sampling. Blood samples were sent to the Pediatric research laboratory, Linköping University, at room-temperature and were processed within 27 h after sample collection. Plasma was separated from heparinized blood and stored at −70°C prior to analysis. PBMC were isolated by performing density-gradient centrifugation with Leucosep® tubes (Greiner Bio One; Austria) and Ficoll-Paque™ PLUS (density 1.077 ± 0.001 g/mL, GE Healthcare; UK) according to manufacturer's instructions. Cells were washed twice in RPMI 1640 w/o L-glutamine (Gibco; US) supplemented with 2 % (v/v) heat-inactivated fetal calf serum (Gibco; US).

Peripheral venous blood was collected in three Vacuette® sodium-heparin tubes à 9 mL (Greiner Bio-One; Austria) in the morning (8–10 a.m.). All study subjects had been fasting since 10 p.m. the night before sampling. Blood samples were sent to the Pediatric research laboratory, Linköping University, at room-temperature and were processed within 27 h after sample collection. PBMC were obtained by performing density-gradient centrifugation with Leucosep® tubes (Greiner Bio One; Austria) and Ficoll-Paque™ PLUS (density 1.077 ± 0.001 g/mL, GE Healthcare; UK) according to manufacturer's instructions. Cells were washed in RPMI 1640 w/o L-glutamine (Gibco; US) supplemented with 2% (v/v) heat-inactivated FCS (Gibco; US). PBMC were incubated over-night in complete medium (RPMI 1640 w/o L-glutamine supplemented with 10% (v/v) heat-inactivated FCS) at 2 × 10⁶ cells/mL.