Vacutainer lh

Biological standard follow-up included sampling of dipotassium EDTA tubes (BD Vacutainer EDTA, Plymouth) for whole blood counts and plasma from lithium heparin tubes with gel separator (BD Vacutainer LH, Plymouth) for biochemical parameters.
The severity of haemolysis was estimated using plasma lactate dehydrogenase level (LDH). These tests were performed in samples collected for routine follow-up. Whole blood counts were measured on XN-9000 (Sysmex, Villepinte, France). LDH, creatinine, ferritin and indirect bilirubin levels were determined on cobas^® 8000 chemistry analyser (Roche Diagnostics, Mannheim, Germany).

Peripheral blood was collected into BD Vacutainer SST II Advance and BD Vacutainer LH (both manufactured by BD Vacutainer Systems, Plymouth, UK). Following centrifugation, serum was aliquoted and stored at −80 °C. PBMCs were isolated from heparinised blood by density centrifugation (Lymphoprep, Axis-Shield, Oslo, Norway) and either analysed immediately (B cells) or cryopreserved in foetal bovine serum with 10% dimethyl sulphoxide (both from Sigma-Aldrich, St Louis, MO, USA) for later analysis (T cells). Prior to use, frozen PBMCs were thawed following the ‘warm’ and ‘slow’ methodology,^{35 (link)} using HyClone Roswell Park Memorial Institute 1640 medium (GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% foetal bovine serum.

Ten mL of venous blood from the antecubital vein was collected on day 1 and 24, 27, 36, and 48 hours following ECs. Venous blood was collected into lithium heparin-containing tubes (BD vacutainer LH; BD Diagnostics, Nu-Care, Bedfordshire, UK) and centrifuged immediately at 3000 x g at 4°C for 10 minutes. Blood plasma was aliquoted and frozen at -80^oC for subsequent analysis. Hydrogen isotope ratios (²H/¹H) of plasma were determined in triplicate by injecting samples into a high-temperature conversion elemental analyzer (TCEA Flash 2000; ThermoFisher Scientific, Waltham, MA, USA) coupled to an isotope ratio mass spectrometer ([IRMS] Delta V; ThermoFisher Scientific). Raw isotope ratio values were normalized with in-house reference materials calibrated to Vienna Standard Mean Ocean Water.