title: Comparison of the effects of glycerol, dimethyl sulfoxide, and hydroxyethyl starch solutions for cryopreservation of avian red blood cells.
abstract: OBJECTIVE
  To compare effectiveness of glycerol, dimethyl sulfoxide (DMSO), and hydroxyethyl starch (HES) solutions for cryopreservation of avian RBCs.
  
  
  METHODS
  RBCs from 12 healthy Ameraucana hens (Gallus gallus domesticus).
  
  
  METHODS
  RBCs were stored in 20% (wt/vol) glycerol, 10% (wt/vol) DMSO freezing medium, or various concentrations of HES solution (7.5%, 11.5%, and 20% [wt/vol]) and frozen for 2 months in liquid nitrogen. Cells were then thawed and evaluated by use of cell recovery and saline stability tests, cell staining (7-aminoactinomycin D and annexin V) and flow cytometry, and scanning electron microscopy.
  
  
  RESULTS
  Percentage of RBCs recovered was highest for 20% glycerol solution (mean ± SE, 99.71 ± 0.04%) and did not differ significantly from the value for 7.5% HES solution (99.57 ± 0.04%). Mean saline stability of RBCs was highest for 10% DMSO (96.11 ± 0.25%) and did not differ significantly from the value for 20% HES solution (95.74 ± 0.25%). Percentages of cells with 7-aminoactinomycin D staining but without annexin V staining (indicating necrosis or late apoptosis) were lowest for 10% DMSO freezing medium (3%) and 20% glycerol solution (1%) and highest for all HES concentrations (60% to 80%). Scanning electron microscopy revealed severe membrane changes in RBCs cryopreserved in 20% HES solution, compared with membrane appearance in freshly harvested RBCs and RBCs cryopreserved in 10% DMSO freezing medium.
  
  
  CONCLUSIONS
  Cryopreservation of avian RBCs with HES solution, regardless of HES concentration, resulted in greater degrees of apoptosis and cell death than did cryopreservation with other media. Transfusion with RBCs cryopreserved in HES solution may result in posttransfusion hemolysis in birds.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Graham
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Meola
authors.forename: Nisha R
authors.initials: NR
authors.lastname: Kini
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Hoffman
mesh_terms: D000818:Animals; D001717:Birds / Q000097:blood*; D002645:Chickens / Q000097:blood*; D015925:Cryopreservation / Q000662:veterinary; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004906:Erythrocyte Count / Q000662:veterinary; D004912:Erythrocytes / Q000187:drug effects*; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D006895:Hydroxyethyl Starch Derivatives / Q000494:pharmacology
chemical_list: D003451:Cryoprotective Agents; D006895:Hydroxyethyl Starch Derivatives; D005990:Glycerol; D004121:Dimethyl Sulfoxide