Novel method to leukoreduce murine blood for transfusion: How to reduce animal usage

Abstract

BACKGROUND Basic research on the pathomechanisms of transfusion-related adverse events depends on murine transfusion models, in which leukoreduction (LR) is a prevalent standard. The commonly used neonatal LR filter (LRF) is associated with considerable animal numbers. A more efficient method would help support the guiding principles of "replacement, reduction, refinement" (3Rs). STUDY DESIGN AND METHODS Blood from C57BL/6 and C57BL/6-Tg(UBC-GFP)30Scha/J mice was leukoreduced using 1) a neonatal LRF, 2) a syringe LRF, or 3) CD45 microbeads. Product quality was assessed according to US Food and Drug Administration (FDA) standards. White blood cell numbers were analyzed by flow cytometry; hemoglobin concentrations and hematocrit were measured and in vivo posttransfusion recoveries were determined after 2 weeks of storage. RESULTS Using the neonatal filter, a LR of 99.56% was achieved with wastage of 12.4 mL in comparison to 99.68% and 1-mL hold-up volume with the syringe filter and 99.11 ± 0.24% LR and 0.1-mL wastage using microbeads. All techniques achieved FDA quality standards, apart from posttransfusion recovery rate, which was only reached by the microbeads-based technique. CONCLUSION LR with CD45 microbeads not only reduces animal usage but also provides a more efficacious method regarding posttransfusion red blood cell recovery and, hence, provides a promising alternative to commonly used methods.