Red cell storage for 56 days in modified DPD-adenine. An in vitro evaluation.

The modified CPD-adenine anticoagulants CPDA-2 and CPDA-3 were developed to improve red blood cell storage to 35 days, since CPDA-1 was found marginal at 35 days in high hematocrit samples. In this study red blood cell storage was extended to 56 days. In vitro correlates of viability were monitored to determine the feasibility of evaluating in vivo the ability of CPDA-2 and CPDA-3 to extend storage past 35 days. The data suggest that red blood cells stored up to 56 days may have acceptable viability, providing the possibility of extended storage for the military and certain special civilian situations.

Improved red blood cell storage using optional additive systems (OAS) containing adenine, glucose and ascorbate-2-phosphate.

Red blood cells were treated with optional additive system (OAS) solutions to provide component-specific metabolic enhancement for improved storage. Red blood cell viability, as monitored by ATP concentrations, was maintained by use of adenine and extra glucose. Red blood cell oxygen offloading characteristics were improved by maintenance of red blood cell 2,3-DPG concentrations with ascorbate-2-phosphate (AsP). The use of CPD-collected red blood cells with an OAS containing adenine, glucose, and AsP, or CPD-adenine collected red blood cells with an OAS containing AsP demonstrates the potential to store red blood cells at least 42 days and to maintain red blood cell 2,3-DPG.

Red cell ATP and 2,3-diphosphoglycerate concentrations as a function of dihydroxyacetone supplementation of CPD adenine.

Units of CPDA-1 whole blood were subdivided and each treated with additions of dihydroxyacetone (DHA) to give final concentrations from 0 to 80 mM. The 'optimum' concentration of DHA to maintain 2,3-diphosphoglycerate (2,3-DPG) with minimal loss of ATP during 42 days of storage appeared to be 30 mM of DHA. With this formulation, red cell 2,3-DPG concentrations rose to 130-140% of normal by 14 days and then decreased in a near-linear manner to 50-60% normal by 42 days, while maintaining adequate ATP levels. In addition, packed red cells were prepared form CPD fresh blood and treated with adenine, glucose, and various concentrations (0-80 mM) of DHA. The cells also responded most favorably to 30mM DHA, although the response was not as positive as whole blood. This concentration of DHA produced nearly 100% maintenance of 2,3-DPG at 14 days with subsequent fall to 30% of normal by 42 days.

The potential use of dihydroxyacetone for improved 2,3-DPG maintenance in red blood cell storage: solution stability and use in packed cell storage.

Dihydroxyacetone (DHA) is effective in maintaining 2,3-diphosphoglycerate (2,3-DPG) concentrations in stored red blood cells. One limitation to the use of DHA is its instability when added to anticoagulant solutions during blood bag manufacture. The stability of DHA solutions have been evaluated. Solutions of DHA are stable at 25 C in water or isotonic saline, with or without the addition of glucose or adenine. DHA is stable to autoclaving; 99 + per cent surviving at 150 mM, and 89 per cent surviving at 1.9 M concentrations. DHA can be incorporated into a satellite addition pouch attached to the main blood drawing bag, and be added to the blood-anticoagulant mixture after phlebotomy or the preparation of red blood cells. Addition of the DHA solution, containing adenine and extra glucose, to packed cells causes significantly improved maintenance of 2,3-DPG during 42 days of 4 C storage, while maintaining adequate concentrations of red blood cell ATP. The use of DHA, adenine, and glucose in extended storage of packed cells, using either zero or seven day addition of the nutrient solution, produces similar efficacious results.