Rejuvenation of RBCs: validation of a manufacturing method suitable for clinical use

Area of Science:

• Blood banking and transfusion medicine
• Cellular metabolism and preservation
• Clinical trial manufacturing

Background:

• Stored red blood cells (RBCs) lose adenosine 5'-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG), impacting oxygen delivery.
• RBC rejuvenation aims to restore these critical metabolic components to levels found in fresh cells.
• Optimizing and validating a US-approved rejuvenation process for a UK clinical trial setting is essential.

Purpose of the Study:

• To optimize and validate the US-approved RBC rejuvenation process for use in a UK multicenter clinical trial.
• To assess the efficacy of rejuvenation in restoring ATP and 2,3-DPG levels in stored RBCs.
• To ensure the safety and quality of rejuvenated RBCs for transfusion in cardiac surgery patients.

Main Methods:

• Leukoreduced RBC units were rejuvenated using a solution of pyruvate, inosine, phosphate, and adenine (Rejuvesol).
• Laboratory studies measured key parameters including ATP, 2,3-DPG, hemolysis, and potassium over 96 hours post-rejuvenation.
• Operational validation involved two phases, with results from the first phase informing specifications for the second, including assessment of crossmatch reactivity and antigen profiles.

Main Results:

• Rejuvenation successfully increased 2,3-DPG and ATP levels to those of fresh RBCs.
• The final rejuvenated product exhibited lower potassium and hemolysis levels compared to standard storage.
• The process did not adversely affect RBC crossmatch reactivity or antigen profiles, meeting trial specifications.

Conclusions:

• The validated RBC rejuvenation process operates within defined quality limits.
• This process effectively preserves RBC immunophenotypes and metabolic function.
• Manufacture of rejuvenated RBCs is feasible for clinical trials, ensuring component quality.