title: Stored red blood cell susceptibility to in vitro transfusion-associated stress conditions is higher after longer storage and increased by storage in saline-adenine-glucose-mannitol compared to AS-1.
abstract: BACKGROUND
  Biochemical changes induced in red blood cells (RBCs) during storage may impair their function upon transfusion. Transfusion-associated stresses may further amplify storage lesion effects including increased phosphatidylserine (PS) exposure at the RBC membrane, microparticle (MP) release, and adhesion to endothelial cells (ECs). RBC stress susceptibility in vitro was investigated in relation to storage time and additive solution.
  
  
  METHODS
  Leukoreduced whole blood donations (n = 18) were paired, mixed, and resplit before separating the RBCs for storage in saline-adenine-glucose-mannitol (SAGM) or AS-1. Samples were taken after 3, 21, or 35 days. For oxidative stress treatment, RBCs were exposed to 0.5 mmol/L tert-butylhydroperoxide. Transfusion-associated stress was simulated by overnight culture at 37 °C with plasma containing inflammatory mediators. PS exposure and MPs were measured by flow cytometry and adhesion to ECs was tested under flow conditions. PS specificity of adhesion was tested by blocking with PS-containing lipid vesicles.
  
  
  RESULTS
  Oxidative stress induced significantly higher PS exposure and adhesion to ECs in RBCs stored for 35 days compared to 3 days (p < 0.04). PS-containing vesicles blocked RBC-EC adhesion. After overnight culture with or without plasma, PS exposure and EC adhesion were significantly increased (p < 0.05). MP numbers increased with longer RBC storage and after RBC culture with plasma. Culture conditions influenced MP numbers from Day 35 RBCs. RBCs stored in SAGM had significantly higher PS exposure after stress treatment than AS-1 RBCs (p < 0.02).
  
  
  CONCLUSIONS
  Storage for 35 days significantly increased RBC susceptibility to oxidative and in vitro transfusion-associated stresses and was higher for RBCs stored in SAGM compared to AS-1.
authors.affiliation: formerly Research and Development, Australian Red Cross Blood Service, West Melbourne, Victoria, Australia.
authors.forename: Diana
authors.initials: D
authors.lastname: Mittag
authors.affiliation: formerly Research and Development, Australian Red Cross Blood Service, West Melbourne, Victoria, Australia.
authors.forename: Amrita
authors.initials: A
authors.lastname: Sran
authors.affiliation: formerly Research and Development, Australian Red Cross Blood Service, West Melbourne, Victoria, Australia.
authors.forename: Kasey S
authors.initials: KS
authors.lastname: Chan
authors.affiliation: School of Psychological and Clinical Sciences, Charles Darwin University, Darwin, Northern Territory, Australia.
authors.forename: Martin P
authors.initials: MP
authors.lastname: Boland
authors.affiliation: Department of Medical Biology, Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
authors.forename: Esther
authors.initials: E
authors.lastname: Bandala-Sanchez
authors.affiliation: Baker IDI Heart and Diabetes Institute, Melbourne, Victoria.
authors.forename: Olivier
authors.initials: O
authors.lastname: Huet
authors.affiliation: formerly Research and Development, Australian Red Cross Blood Service, West Melbourne, Victoria, Australia.
authors.forename: William
authors.initials: W
authors.lastname: Xu
authors.affiliation: formerly Research and Development, Australian Red Cross Blood Service, West Melbourne, Victoria, Australia.|Department of Immunology, Monash University, Melbourne, Victoria, Australia.
authors.forename: Rosemary L
authors.initials: RL
authors.lastname: Sparrow
mesh_terms: D000225:Adenine / Q000494:pharmacology*; D001793:Blood Preservation; D002448:Cell Adhesion / Q000187:drug effects; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism*; D017707:Erythrocyte Transfusion; D005260:Female; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D008297:Male; D008353:Mannitol / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects*; D019999:Pharmaceutical Solutions / Q000494:pharmacology; D010718:Phosphatidylserines / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology*; D013997:Time Factors
chemical_list: C037839:ADSOL; D019999:Pharmaceutical Solutions; D010718:Phosphatidylserines; D008353:Mannitol; D012965:Sodium Chloride; D005947:Glucose; D000225:Adenine