Use of an in vitro biotinylation technique for determination of posttransfusion survival of fresh and stored autologous red blood cells in Thoroughbreds.
Abstract: 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.
Publication Date: 2010-08-03 PubMed ID: 20673097DOI: 10.2460/ajvr.71.8.960Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article is a study on the use of a biotinylation technique to determine the survival rate of both fresh and preserved red blood cells in Thoroughbred horses after transfusion.
Research Methodology
- The researchers evaluated the use of N-hydroxysuccinimide (NHS)-biotin — a tracer compound — to label red blood cells (RBCs) in Thoroughbred horses.
- The study involved serial dilutions of biotin and streptavidin-phycoerythrin (PE) on blood samples from three horses to perfect the procedure in vitro.
- The researchers subsequently moved to in vivo experiments involving another twelve horses, divided into four groups.
- Blood was collected from these horses into a solution of citrate phosphate dextrose adenine-1 (CPDA-1) — a commonly used blood preservative — and then labelled with biotin. This blood was then either immediately reinfused to the horses or stored at 4 degrees Celsius for 1, 14, or 28 days prior to reinfusion.
Observations & Results
- From their in vitro studies, the researchers were able to determine optimal concentrations for biotin and streptavidin-PE in this application.
- The post-transfusion survival of RBCs — meaning how long these cells continued to function — was monitored and recorded at various intervals for 35 days after reinfusion.
- The results of this monitoring indicated that the lifespan of RBCs after transfusion was 99, 89, 66, and 59 days after storage for 0, 1, 14, and 28 days respectively. This showed that storage did not result in drastic alterations in RBC lifespan.
- The average half-life — the time it took for half of the RBCs to cease functioning post-transfusion — was 50, 45, 33, and 29 days for 0, 1, 14, and 28 days of storage respectively.
Conclusions
- The study concluded that NHS-biotin can be effectively used for labelling RBCs in horses and provides a means to monitor their survival post-transfusion.
- The survival rates of RBCs post-transfusion that they observed were higher than those previously reported.
Cite This Article
APA
Owens SD, Johns JL, Walker NJ, Librach FA, Carrade DD, Tablin F, Borjesson DL.
(2010).
Use of an in vitro biotinylation technique for determination of posttransfusion survival of fresh and stored autologous red blood cells in Thoroughbreds.
Am J Vet Res, 71(8), 960-966.
https://doi.org/10.2460/ajvr.71.8.960 Publication
Researcher Affiliations
- Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616, USA. sdowens@ucdavis.edu
MeSH Terms
- Animals
- Biotinylation / methods
- Biotinylation / veterinary
- Cell Survival / physiology
- Erythrocyte Transfusion / methods
- Erythrocyte Transfusion / veterinary
- Erythrocytes / cytology
- Erythrocytes / drug effects
- Half-Life
- Horses / blood
- Regression Analysis
Citations
This article has been cited 1 times.- Tomlinson JE, Taberner E, Boston RC, Owens SD, Nolen-Walston RD. Survival Time of Cross-Match Incompatible Red Blood Cells in Adult Horses. J Vet Intern Med 2015 Nov-Dec;29(6):1683-8.
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