Peter A Smethurst1, Jennifer Jolley1, Rebecca Braund2, Sue Proffitt1, Thomas Lynes3, Matthew Hazell3, Phil Mellor2, Kay Ridgwell4, Simon Procter5, Alexandra Griffiths6, Anthony M Marinaki7, Helen V New8,9, Gavin J Murphy10, Dave Edmondson2, Rebecca Cardigan1,11. 1. Component Development Laboratory, NHS Blood and Transplant, Cambridge, United Kingdom. 2. Manufacturing & Development, NHS Blood and Transplant, Bristol, United Kingdom. 3. Red Cell Immunohaematology, NHS Blood and Transplant, Bristol, United Kingdom. 4. IBGRL Protein Development & Production Unit, NHS Blood and Transplant, Bristol, United Kingdom. 5. Quality Monitoring, NHS Blood and Transplant, London, United Kingdom. 6. Clinical Trials Unit, NHS Blood and Transplant, Bristol, United Kingdom. 7. Purine Research Laboratory, St Thomas' Hospital, London, United Kingdom. 8. Clinical Directorate, NHS Blood and Transplant, London, United Kingdom. 9. Department of Haematology, Imperial College London, London, United Kingdom. 10. Department of Cardiovascular Sciences and NIHR Leicester Biomedical Research Centre, Glenfield General Hospital, University of Leicester, Leicester, United Kingdom. 11. Department of Haematology, University of Cambridge, Cambridge, United Kingdom.
Abstract
BACKGROUND: Rejuvenation of stored red blood cells (RBCs) increases levels of adenosine 5'-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) to those of fresh cells. This study aimed to optimize and validate the US-approved process to a UK setting for manufacture and issue of rejuvenated RBCs for a multicenter randomized controlled clinical trial in cardiac surgery. STUDY DESIGN AND METHODS: Rejuvenation of leukoreduced RBC units involved adding a solution containing pyruvate, inosine, phosphate, and adenine (Rejuvesol, Zimmer Biomet), warming at 37°C for 60 minutes, then "manual" washing with saline adenine glucose mannitol solution. A laboratory study was conducted on six pools of ABO/D-matched units made the day after donation. On Days 7, 21, and 28 of 4 ± 2°C storage, one unit per pool was rejuvenated and measured over 96 hours for volume, hematocrit, hemolysis, ATP, 2,3-DPG, supernatant potassium, lactate, and purines added (inosine) or produced (hypoxanthine) by rejuvenation. Subsequently, an operational validation (two phases of 32 units each) was undertaken, with results from the first informing a trial component specification applied to the second. Rejuvenation effects were also tested on crossmatch reactivity and RBC antigen profiles. RESULTS: Rejuvenation raised 2,3-DPG to, and ATP above, levels of fresh cells. The final component had potassium and hemolysis values below those of standard storage Days 7 and 21, respectively, containing 1.2% exogenous inosine and 500 to 1900 μmoles/unit of hypoxanthine. The second operational validation met compliance to the trial component specification. Rejuvenation did not adversely affect crossmatch reactivity or RBC antigen profiles. CONCLUSION: The validated rejuvenation process operates within defined quality limits, preserving RBC immunophenotypes, enabling manufacture for clinical trials.
BACKGROUND: Rejuvenation of stored red blood cells (RBCs) increases levels of adenosine 5'-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) to those of fresh cells. This study aimed to optimize and validate the US-approved process to a UK setting for manufacture and issue of rejuvenated RBCs for a multicenter randomized controlled clinical trial in cardiac surgery. STUDY DESIGN AND METHODS: Rejuvenation of leukoreduced RBC units involved adding a solution containing pyruvate, inosine, phosphate, and adenine (Rejuvesol, Zimmer Biomet), warming at 37°C for 60 minutes, then "manual" washing with saline adenine glucose mannitol solution. A laboratory study was conducted on six pools of ABO/D-matched units made the day after donation. On Days 7, 21, and 28 of 4 ± 2°C storage, one unit per pool was rejuvenated and measured over 96 hours for volume, hematocrit, hemolysis, ATP, 2,3-DPG, supernatant potassium, lactate, and purines added (inosine) or produced (hypoxanthine) by rejuvenation. Subsequently, an operational validation (two phases of 32 units each) was undertaken, with results from the first informing a trial component specification applied to the second. Rejuvenation effects were also tested on crossmatch reactivity and RBC antigen profiles. RESULTS: Rejuvenation raised 2,3-DPG to, and ATP above, levels of fresh cells. The final component had potassium and hemolysis values below those of standard storage Days 7 and 21, respectively, containing 1.2% exogenous inosine and 500 to 1900 μmoles/unit of hypoxanthine. The second operational validation met compliance to the trial component specification. Rejuvenation did not adversely affect crossmatch reactivity or RBC antigen profiles. CONCLUSION: The validated rejuvenation process operates within defined quality limits, preserving RBC immunophenotypes, enabling manufacture for clinical trials.
Authors: Aline Roch; Nicholas J Magon; Jessica Maire; Cacang Suarna; Anita Ayer; Sophie Waldvogel; Beat A Imhof; Mark J Koury; Roland Stocker; Marc Schapira Journal: JCI Insight Date: 2019-11-01