Abstract
Background - The quality of red blood cells (RBCs) stored in red cell concentrates (RCCs) is influenced by processing, storage and donor characteristics, and can have a clinical impact on transfused patients.To evaluate RBC properties and their potential impact in a transfusion setting, a simple in vitro-transfusional model has been developed.
Materials and methods - Transfusion was simulated by mixing a washed RBC pool from two male-derived RCCs stored at 4°C with a pool of 15 male-derived fresh frozen plasma (FFP) units, representing the recipient, at a hematocrit (HCT) of 30% (“control” setting) or 5% (alternative model).The mixtures were incubated at 37°C, 5% of CO2 up to 48 h.Different metabolites, hemolysis and microvesicles (MVs) were quantified at several incubation times and RBC-morphology changes and deformability after incubation. For each model, biological triplicates have been investigated with RCCs at storage days 2 and 43.
Results - The 5%-HCT model restored the 2,3-DPG level and maintained the ATP level. Furthermore, glucose consumption and corresponding lactate production were increased in the 5%- vs the 30%-HCT condition. Lower hemolysis was observed with 5%-HCT, but only at day 2.However, morphological analysis by digital holographic microscopy (DHM) revealed a decreased fraction of discocytes at 5% rather than at 30% of HCT at storage day 2 but at day 43, the trend was inverted. Concordantly, RBCs incubated at 5% of HCT were more deformable than at 30% at day 43 (p<0.0001).
Discussion - Higher metabolic activity of RBCs in the 5%-HCT condition was promoted by a higher glucose availability and limited cell-waste accumulation.The conditions of the new proposed model thus enabled rejuvenation of RBCs and maintained them in a physiological-close state in contrast to the 30%-HCT model.It may be used as a first approach to evaluate e.g., the impact of donor and recipient characteristics on RBC properties.
References
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Authors
Emmanuel Längst Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interrégionale CRS SA, Epalinges, Switzerland; Faculté de Biologie et de Médecine, University of Lausanne (UNIL), Lausanne, Switzerland
David Crettaz Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interrégionale CRS SA, Epalinges, Switzerland
Julien Delobel
Laboratoire de Recherche and Unité d’Hématologie-Oncologie Pédiatrique, Service de Pédiatrie, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland
Raffaele Renella Laboratoire de Recherche and Unité d’Hématologie-Oncologie Pédiatrique, Service de Pédiatrie, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland
Manon Bardyn
Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interrégionale CRS SA, Epalinges, Switzerland
Gerardo Turcatti
Biomolecular Screening Facility (BSF), Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland
Jean-Daniel Tissot
Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interrégionale CRS SA, Epalinges, Switzerland
Michel Prudent
Laboratoire de Recherche sur les Produits Sanguins, Transfusion Interrégionale CRS SA, Epalinges, Switzerland; Faculté de Biologie et de Médecine, University of Lausanne (UNIL), Lausanne, Switzerland; Center for Research and Innovation in Clinical Pharmaceutical Sciences, Institute of Pharmaceutical Sciences of Western Switzerland, Centre Hospitalier Universitaire Vaudois (CHUV) and University of Lausanne (UNIL), Lausanne, Switzerland; Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva (UNIGE), University of Lausanne (UNIL), Switzerland
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