Alves Diana, Sparrow Rosemary, Garnier Gil
Bioresource Processing Research Institute of Australia (BioPRIA), Department of Chemical Engineering, Monash University, Clayton, Victoria, Australia.
Transfusion Research Unit, Department of Epidemiology and Preventive Medicine, School of Public Health and Preventive Medicine, Monash University, Melbourne, Victoria, 3004, Australia.
J Biomed Mater Res B Appl Biomater. 2021 Nov;109(11):1689-1697. doi: 10.1002/jbm.b.34825. Epub 2021 Mar 10.
Prior to transfusion of red blood cells (RBCs), recipients must be tested for the presence of alloantibodies to avoid immune complications. Liquid-preserved reagent RBCs with known blood group antigen phenotypes are used for testing. However, these reagents have practical constraints, including limited shelf-life and require constant refrigeration. To address these issues, we explore the effects of rapid freeze-drying conditions with trehalose cryoprotectant (0.1-1 M concentrations) on human RBCs and storage of freeze-dried RBCs (FDRBCs) at room temperature (RT) for up to 12 months. We report that rapid freeze-drying of RBCs for 2.5 hr with 0.5 M trehalose achieves recoverable cells with near-normal morphological shape, although size-reduced. The FDRBCs are metabolically active and functional in antibody-agglutination tests by the column agglutination test (CAT) for ABO and Rhesus-D blood group antigens. Expression of the Duffy blood group protein (CD234) decreases by 50% after freeze-drying RBCs. The initial recovery rate is ≤25%; however, 43% of these FDRBCs are still recoverable after RT storage for 12 months. In this proof-of-principle study, we show that rapid freeze-drying can stabilize RBCs. Further refinements to improve the recovery rate and preservation of antigenic epitopes will make FDRBCs a practical alternative source of reagent RBCs for pre-transfusion alloantibody identification.
在输注红细胞（RBC）之前，必须对受血者进行同种抗体检测，以避免免疫并发症。使用具有已知血型抗原表型的液体保存试剂红细胞进行检测。然而，这些试剂存在实际限制，包括保质期有限且需要持续冷藏。为了解决这些问题，我们探究了使用海藻糖冷冻保护剂（浓度为0.1 - 1M）的快速冻干条件对人红细胞的影响，以及冻干红细胞（FDRBC）在室温（RT）下长达12个月的储存情况。我们报告称，用0.5M海藻糖对红细胞进行2.5小时的快速冻干可获得形态接近正常但尺寸减小的可恢复细胞。通过柱凝集试验（CAT）检测ABO和恒河猴-D血型抗原，FDRBC在抗体凝集试验中具有代谢活性且功能正常。冻干红细胞后，达菲血型蛋白（CD234）的表达降低50%。初始恢复率≤25%；然而，在室温下储存12个月后，这些FDRBC中有43%仍可恢复。在这项原理验证研究中，我们表明快速冻干可使红细胞稳定。进一步改进以提高恢复率并保留抗原表位将使FDRBC成为输血前同种抗体鉴定中试剂红细胞的一种实用替代来源。
