One of the most common life-saving medical procedures is a red blood cell (RBC) transfusion. Unfortunately, RBCs for transfusion have a limited shelf life after donation due to detrimental storage effects on their morphological and biochemical properties. Inspired by nature, a biomimetics approach was developed to preserve RBCs for long-term storage using compounds found in animals with a natural propensity to survive in a frozen or desiccated state for decades. Trehalose was employed as a cryoprotective agent and added to the extracellular freezing solution of porcine RBCs. Slow cooling (-1 C min-1) resulted in almost complete hemolysis (1 ± 1 % RBC recovery), and rapid cooling rates had to be used to achieve satisfactory cryopreservation outcomes. After rapid cooling, the highest percentage of RBC recovery was obtained by plunging in liquid nitrogen and thawing at 55 C, using a cryopreservation solution containing 300 mM trehalose. Under these conditions, 88 ± 8 % of processed RBCs were recovered and retained hemoglobin (14 ± 2 % hemolysis). Hemoglobin’s oxygen-binding properties of cryopreserved RBCs were not significantly different to unfrozen controls and was allosterically regulated by 2,3-bisphosphoglycerate. These data indicate the feasibility of using trehalose instead of glycerol as a cryoprotective compound for RBCs. In contrast to glycerol, trehalose-preserved RBCs can potentially be transfused without time-consuming washing steps, which significantly facilitates the usage of cryopreserved transfusible units in trauma situations when time is of the essence.
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This content will become publicly available on January 1, 2025
Rehydration outcomes for freeze-dried red blood cells in reduced gravity
Medical planning for space exploration is based on the “floating” blood bank model to store life-saving red blood cells (RBCs) for emergencies. The “floating” blood bank approach is not sufficient in cases where multiple crewmembers are affected by space anemia. In these situations, long-term preserved RBCs will be vital to guarantee the health and safety of crew members. Transfusable RBC units can only be refrigerated for 42 days or frozen at -80 C. However, storing frozen RBCs at -80 C is challenging during the confined condition of long-duration space flight. Freeze-dried, viable RBCs would be an appropriate alternative because they can be stored without cooling, are predicted to have a shelf-life of years, and could be transfused immediately after rehydration. This study explores if freeze-dried RBCs can be rehydrated and transfused in reduced gravity with similar outcomes in recovery as observed at Earth gravity. Experiments analyzing freeze-dried RBC recoveries, rehydration fluid dynamics, and transfusion flow rates were analyzed utilizing an experimental glovebox in simulated 0 g during parabolic flights. RBC recoveries and rehydration fluid dynamics for volumes of 5 mL and 10 mL were the same in simulated 0 g compared to results obtained at 1 g. A clinically acceptable range of flow rates for slow intravenous infusion and rapid fluid resuscitation was possible with the simple augmentation of a hand-pumped clinical pressure bag around a unit of rehydrated RBCs. The results demonstrate the potential feasibility of using freeze-dried cells for healthcare during deep-space exploration.
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- Award ID(s):
- 1827521
- NSF-PAR ID:
- 10478726
- Publisher / Repository:
- Elsevier
- Date Published:
- Journal Name:
- Acta Astronautica
- Volume:
- 214
- Issue:
- C
- ISSN:
- 0094-5765
- Page Range / eLocation ID:
- 64 to 71
- Subject(s) / Keyword(s):
- Parabolic flight freeze-dried RBCs 0 g fluid dynamics transfusion anhydrobiosis
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Methods Atomic force microscopy (AFM) was used to assess stiffness of RBCs after exposure to different IVFs. A microfluidic model of the human capillary system was used to assess transit time (TT) and propensity to occlusion after exposure to the different IVF formulations.
Results Sickle RBCs exposed to normal saline (NS) had increased stiffness, TTs, and propensity to microchannel occlusion compared to other osmolarities.
Conclusion NS, an IVF formulation often used to treat patients with SCD during VOC, may induce localized microvascular obstruction due to alterations of sickle RBC biomechanical properties.
