Use of an in vitro biotinylation technique for determination of posttransfusion survival of fresh and stored autologous red blood cells in Thoroughbreds.
OBJECTIVE
To evaluate N-hydroxysuccinimide (NHS)-biotin labeling of equine RBCs and determine posttransfusion survival of autologous equine RBCs stored in citrate phosphate dextrose adenine-1 (CPDA-1) for 0, 1, 14, and 28 days.


METHODS
13 healthy adult Thoroughbreds.


METHODS
Serial dilutions of biotin and streptavidin-phycoerythrin (PE) were evaluated in vitro in blood collected from 3 horses. One horse was used to determine RBC distribution and recovery. Twelve horses were allocated to 4 groups for in vivo experiments in which blood was collected into CPDA-1. Blood was labeled with biotin and reinfused or stored at 4 degrees C for 1, 14, or 28 days prior to labeling with NHS-biotin and reinfusion. Posttransfusion blood samples were collected 15 minutes and 1, 2, 3, 5, 7, 14, 21, 28, and 35 days after reinfusion. Biotin-labeled RBCs were detected via flow cytometry by use of streptavidin-PE. Posttransfusion lifespan of RBCs and RBC half-life were determined.


RESULTS
Optimal biotin concentration was 0.04 pg of biotin/RBC, and the optimal streptavidin-PE ratio was 1.2 microg of streptavidin-PE/1 x 10(6) RBCs. Posttransfusion lifespan of autologous RBCs was 99, 89, 66, and 59 days after storage for 0, 1, 14, and 28 days, respectively. Storage did not result in significant alterations in RBC lifespan. Mean posttransfusion RBC half-life was 50, 45, 33, and 29 days for 0, 1, 14, and 28 days of storage, respectively.


CONCLUSIONS
Biotin can be used to label equine RBCs for RBC survival studies. Posttransfusion survival of equine autologous RBCs was greater than previously reported.
Owens
Sean D
SD
Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616, USA. sdowens@ucdavis.edu
Johns
Jennifer L
JL
Walker
Naomi J
NJ
Librach
Fred A
FA
Carrade
Danielle D
DD
Tablin
Fern
F
Borjesson
Dori L
DL
D000818:Animals; D019921:Biotinylation / Q000379:methods* / Q000662:veterinary; D002470:Cell Survival / Q000502:physiology*; D017707:Erythrocyte Transfusion / Q000379:methods / Q000662:veterinary*; D004912:Erythrocytes / Q000166:cytology* / Q000187:drug effects; D006207:Half-Life; D006736:Horses / Q000097:blood; D012044:Regression Analysis