BACKGROUND: Pathogen reduction technology (PRT) may improve the safety of RBCs for transfusion. As the Czech Republic considers PRT, we asked what effects riboflavin and UV light PRT pre-freezing has on the post-thaw recovery and properties of cryopreserved RBCs (CRBCs) after deglycerolization and liquid storage. STUDY DESIGN AND METHODS: 24 Group O whole blood (WB) units were leukoreduced and then treated with riboflavin and UV light PRT (Mirasol, Terumo BCT, USA) before cryopreservation (T-CRBC); 20 similarly-collected units were untreated controls (C-CRBC). Units were processed to RBCs and then cryopreserved with 40% glycerol (wt/vol), frozen at -80°C, stored >118 days, reconstituted as deglycerolized RBC units in AS-3, and stored at 4 ± 2°C for 21 days. One treated unit sustained massive hemolysis during the post-thaw wash process and was removed from data analysis. The remaining units were assessed pre-PRT, post-PRT, and post-thaw-wash on days 0, 7, 14, and 21 for hematocrit, volume, hemoglobin per transfusion unit, pH, % hemolysis, hemoglobin in the supernatant, potassium, phosphorus, NH3 , osmolality, ATP, and 2,3-diphosphoglycerate. RESULTS: PRT with leukoreduction caused a 5% loss of RBC followed by a 24% freeze-thaw-wash related loss for a total 28% loss but treated units contained an average of 45 g of hemoglobin, meeting European Union guidelines for CRBC. T-CRBCs displayed higher post-wash hemolysis, potassium, and ammonia concentrations, and lower ATP at the end of storage. CONCLUSIONS: Cryopreserved RBCs from Riboflavin and UV light-treated WB meet the criteria for clinical use for 7 days after thawing and provide additional protection against infectious threats.

Department of Anesthesia and Pain Medicine University of Washington Seattle Washington USA

Department of Biochemistry St Sisters of Mercy Hospital Karel Boromejsky Prague Czech Republic

Department of Hematology and Blood Transfusion Military University Hospital Prague Czech Republic

Department of Internal Medicine 4 Hematology University Hospital Hradec Kralove Czech Republic

Department of Laboratory Medicine and Pathology University of Washington Seattle Washington USA

Department of Military Internal Medicine and Military Hygiene Faculty of Military Health Sciences University of Defence Hradec Kralove Czech Republic

Faculty of Biomedical Engineering Czech Technical University Prague Czech Republic

Harborview Injury Prevention and Research Center Harborview Medical Center Seattle Washington USA

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Valeri CR. Blood components in the treatment of acute blood loss: use of freeze-preserved red cells, platelets, and plasma proteins. Anesth Analg. 1975;54(1):1-14.

Hess JR. Red cell freezing and its impact on the supply chain. Transfus Med. 2004;14(1):1-8.

Lagerberg JW. Cryopreservation of red blood cells. Methods Mol Biol. 2015;1257:353-67.

Noorman F, van Dongen TT, Plat MJ, et al. Transfusion: −80°C frozen blood products are safe and effective in military casualty care. PLoS One. 2016;11(12):e0168401.

Peyrard T, Pham BN, Le Pennec PY, Rouger P. Transfusion of rare cryopreserved red blood cell units stored at −80 degrees C: the French experience. Immunohematology. 2009;25(1):13-7.

Ashenden M, Mørkeberg J. Net haemoglobin increase from reinfusion of refrigerated vs. frozen red blood cells after autologous blood transfusions. Vox Sang. 2011;101(4):320-6.

Valeri CR, Ragno G, Pivacek LE, Cassidy GP, Srey R, Hansson-Wicher M, et al. An experiment with glycerol-frozen red blood cells stored at −80 degrees C for up to 37 years. Vox Sang. 2000;79(3):168-74.

Bohonek M, Kutac D, Acker JP, Seghatchian J. Optimizing the supply of whole blood-derived bioproducts through the combined implementation of cryopreservation and pathogen reduction technologies and practices: an overview. Transfus Apher Sci. 2020;59(2):102754.

Prowse CV. Component pathogen inactivation: a critical review. Vox Sang. 2013;104(3):183-99.

Fast LD, DiLeone G, Marschner S. Inactivation of human white blood cells in platelet products after pathogen reduction technology treatment in comparison to gamma irradiation. Transfusion. 2011;51(7):1397-404.

Prowse CV. Pathogen inactivation of blood components. Transfus Alternat Transfus Med. 2008;10:139-46.

Rebulla P. The long and winding road to pathogen reduction of platelets, red blood cells and whole blood. Br J Haematol. 2019;186(5):655-67.

Perez-Pujol S, Tonda R, Lozano M, Fuste B, Lopez-Vilchez I, Galan AM, et al. Effects of a new pathogen-reduction technology (Mirasol PRT) on functional aspects of platelet concentrates. Transfusion. 2005;45(6):911-9.

Mirasol Clinical Evaluation Study Group. A randomized controlled clinical trial evaluating the performance and safety of platelets treated with MIRASOL pathogen reduction technology. Transfusion. 2010;50(11):2362-75.

Capicciotti CJ, Kurach JD, Turner TR, Mancini RS, Acker JP, Ben RN. Small molecule ice recrystallization inhibitors enable freezing of human red blood cells with reduced glycerol concentrations. Sci Rep. 2015;8(5):9692.

D'Alessandro A, Gray AD, Szczepiorkowski ZM, Hansen K, Herschel LH, Dumont LJ. Red blood cell metabolic responses to refrigerated storage, rejuvenation, and frozen storage. Transfusion. 2017;57(4):1019-30.

Valeri CR, Srey R, Tilahun D, Ragno G. The in vitro quality of red blood cells frozen with 40 percent (wt/vol) glycerol at −80 degrees C for 14 years, deglycerolized with the Haemonetics ACP 215, and stored at 4 degrees C in additive solution-1 or additive solution-3 for up to 3 weeks. Transfusion. 2004;44(7):990-5.

Bohonek M, Petras M, Turek I, et al. In vitro parameters of cryopreserved leucodepleted and non-leucodepleted red blood cells collected by apheresis or from whole blood and stored in AS-3 for 21 days after thawing. Blood Transfus. 2014;12(Suppl 1):s199-203.

Bohonek M, Petras M, Turek I, et al. Quality evaluation of frozen apheresis red blood cell storage with 21-day postthaw storage in additive solution 3 and saline-adenine-glucose-mannitol: biochemical and chromium-51 recovery measures. Transfusion. 2010;50(5):1007-1.

Lagerberg JW, Truijens-de Lange R, de Korte D, et al. Altered processing of thawed red cells to improve the in vitro quality during postthaw storage at 4 degrees C. Transfusion. 2007;47(12):2242-9.

Hess JR, Hill HR, Oliver CK, Lippert LE, Greenwalt TJ. The effect of two additive solutions on the postthaw storage of RBCs. Transfusion. 2001;41(7):923-7.

Moore GL, Ledford ME, Mathewson PJ, Hankins DJ, Shah SB. Post-thaw storage at 4 degrees C of previously frozen red cells with retention of 2,3-DPG. Vox Sang. 1987;53(1):15-8.

Decree of the Ministry of Health of Czech Republic No. 143/2008.

Council of Europe. Rec.No.R(20)20 Guide to the preparation, use and quality assurance of blood components.

Standards for blood banks and transfusion services. 32nd ed. Bethesda, Maryland: AABB; 2020.

Valeri CR, Ragno G, Pivacek LE, Srey R, Hess JR, Lippert LE, et al. A multicenter study of in vitro and in vivo values in human RBCs frozen with 40-percent (wt/vol) glycerol and stored after deglycerolization for 15 days at 4 degrees C in AS-3: assessment of RBC processing in the ACP 215. Transfusion. 2001 Jul;41(7):933-9.

Howell A, Turner TR, Hansen A, Lautner LJ, Yi Q, Acker JP. Closed system processing variables affect post-thaw quality characteristics of cryopreserved red cell concentrates. Transfusion. 2022;62:2577-86. https://doi.org/10.1111/trf.17138

Dimberg LY, Doane SK, Yonemura S, Reddy HL, Hovenga N, Gosney EJ, et al. Red blood cells derived from whole blood treated with riboflavin and UV light maintain adequate cell quality through 21 days of storage. Transfus Med Hemother. 2019;46(4):240-7.

Rossi's Principles and Practice of Transfusion Medicine. In: Simon TL et al., editors. Chapter 14, red blood cell metabolism and preservation. Hess JR, D'Alessandro a. Sixth ed. Hoboken, New Jersey, USA: John Wiley & Sons, Ltd; 2002.