S monovette lithium heparin 9 ml tubes

Fresh heparinized blood was collected from healthy pigs in S-Monovette^® Lithium-Heparin 9 mL tubes (Sarstedt, Nümbrecht, Germany). The donor pigs were kept either in the Clinic for Swine, Small Ruminants and Forensic Medicine and Ambulatory Service at the University of Veterinary Medicine Hannover, Foundation, Germany, for sperm donation or in the Research Center for Emerging Infections and Zoonoses from the University of Veterinary Medicine Hannover, Foundation, Germany, as control animals. Pigs included in this study were male and female and were not under treatment, and a general health examination by a trained veterinarian was conducted before the blood donation. The blood was used immediately for neutrophil isolation. Porcine neutrophils were purified using Biocoll (1.077 g/mL, Biochrom, L6115, Berlin, Germany or Biocoll^® (1.077 g/mL, Bio&SELL GmbH, BSL6115, Nürnberg, Germany) and hypotonic lysis of erythrocytes as previously described [51 (link)]. Cells were resuspended in Roswell Park Memorial Institute (RPMI) 1640 without phenol red (Thermo Fisher, 11835063 GibcoTM, Waltham, MA, USA).
Within the assays that show data with the three serotypes, always neutrophils from the same donor pig (for one independent experiment) were used for all serotypes in parallel.

The collection of blood from healthy pigs was registered at the Lower Saxonian State Office for Consumer Protection and Food Safety (Niedersächsisches Landesamt für Verbraucherschutz und Lebensmittelsicherheit, No. 33.9-42502-05-18A302). It was conducted in line with the recommendations of the German Society for Laboratory Animal Science (Gesellschaft für Versuchstierkunde) and the German Veterinary Association for the Protection of Animals (Tierärztliche Vereinigung für Tierschutz e. V.) (http://www.gv-solas.de).
The donor pigs were kept in the Clinic for Swine or the Research Center for Emerging Infections and Zoonoses from the University of Veterinary Medicine Hannover, Germany. A total of 30 mL of fresh blood was collected from healthy pigs in S-Monovette^®® Lithium-Heparin 9 mL tubes (Sarstedt, Nümbrecht Germany). Half of the blood was analyzed immediately (fresh blood, 0 h) and the rest of the blood was stored in the dark at room temperature for 24 h (stored blood, 24 h). Room temperature was chosen to avoid cold shock of the blood cells.
The same assays were performed with fresh and stored blood for a paired analysis.