title: Manufacturing method affects mitochondrial DNA release and extracellular vesicle composition in stored red blood cells.
abstract: OBJECTIVE
  Damage-associated molecular patterns (DAMPs) are found in transfusion products, but their potential impacts are not fully understood. We examined the influence of manufacturing method on levels of mitochondrial (mt) DNA and extracellular vesicle (EV) DAMPs in red cell concentrates (RCCs).
  
  
  METHODS
  Eighty-seven RCCs were prepared using nine different methods (6-15 units/method), including three apheresis, five whole blood (WB)-derived leucoreduced (LR) and one WB-derived non-LR method. On storage days 5 and 42, levels of mtDNA (by PCR) and number and cell of origin of EVs (by flow cytometry) were assessed in RCC supernatants.
  
  
  RESULTS
  There was a 100-fold difference in mtDNA levels among methods, with highest levels in non-LR, followed by MCS+ and Trima apheresis RCCs. There was a 10-fold difference in EV levels among methods. RBC-derived CD235a+ EVs were found in fresh RCCs and increased in most during storage. Platelet-derived CD41a+ EVs were highest in non-LR and Trima RCCs and did not change during storage. WBC-derived EVs were low in most RCCs; CD14+ EVs increased in several RCCs during storage.
  
  
  CONCLUSIONS
  DAMPs in RCCs vary by manufacturing method. MtDNA and EV could be informative quality markers that may be relevant to RCC immunomodulatory potential.
authors.affiliation: Blood Systems Research Institute, San Francisco, CA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Bakkour
authors.affiliation: Canadian Blood Services Centre for Innovation, Edmonton, AB, Canada.|Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, Canada.
authors.forename: J P
authors.initials: JP
authors.lastname: Acker
authors.affiliation: Blood Systems Research Institute, San Francisco, CA, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Chafets
authors.affiliation: Blood Systems Research Institute, San Francisco, CA, USA.
authors.forename: H C
authors.initials: HC
authors.lastname: Inglis
authors.affiliation: Blood Systems Research Institute, San Francisco, CA, USA.|Department of Laboratory Medicine, University of California, San Francisco, CA, USA.|Department of Medicine, University of California, San Francisco, CA, USA.
authors.forename: P J
authors.initials: PJ
authors.lastname: Norris
authors.affiliation: Blood Systems Research Institute, San Francisco, CA, USA.
authors.forename: T-H
authors.initials: TH
authors.lastname: Lee
authors.affiliation: Blood Systems Research Institute, San Francisco, CA, USA.|Department of Laboratory Medicine, University of California, San Francisco, CA, USA.
authors.forename: M P
authors.initials: MP
authors.lastname: Busch
mesh_terms: D001781:Blood Component Removal; D001793:Blood Preservation / Q000379:methods*; D004272:DNA, Mitochondrial / Q000235:genetics / Q000378:metabolism*; D004906:Erythrocyte Count; D004912:Erythrocytes / Q000166:cytology* / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D008928:Mitochondria / Q000235:genetics*; D019007:P-Selectin / Q000378:metabolism; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D016133:Polymerase Chain Reaction; D013997:Time Factors
chemical_list: D004272:DNA, Mitochondrial; D018950:Lipopolysaccharide Receptors; D019007:P-Selectin; D040201:Platelet Membrane Glycoprotein IIb
keywords: blood component; extracellular vesicles; manufacture; microparticles; mitochondrial DNA; process; red blood cells; storage