Thesis
Comparison of established and novel methods for the detection and enumeration of microparticles in canine stored erythrocyte concentrates for transfusion
Washington State University
Master of Science (MS), Washington State University
2015
Handle:
https://hdl.handle.net/2376/102741
Abstract
Microparticles are submicron (< 1[mu]m), phosphatidylserine-bearing lipid vesicles with important physiologic and pathologic roles in coagulation and inflammation. Microparticles accumulate in stored erythrocyte concentrates for transfusion, decreasing the product quality and potentially increasing recipient morbidity. Canine microparticle detection has been based primarily on phosphatidylserine-labeling with annexin V in supernatant samples, but generation of supernatants induces pre-analytical error. In this study, microparticles in canine erythrocyte concentrates were labeled and enumerated at three time points across the erythrocyte concentrate storage period (days 7, 21, and 35) using the standard annexin V along with the novel labels lactadherin, and bio-malemide dye. Annexin V and bio-malemide labeling of microparticles resulted in similar counts at all time points (p = 0.16). Lactadherin labeling resulted in higher microparticle counts than annexin V and bio-malemide at all time points (p = 0.002 and p = 0.006, respectively), particularly on day 7, suggesting non-specific binding. Bio-malemide and annexin V-based microparticle counts increased significantly from day 7 to 35 (p = 0.04) and increases from day 21 to 35 approach significance (p = 0.05). This study indicates that bio-malemide is an alternative to annexin V in canine microparticle research for general microparticle labeling and that in erythrocyte concentrates, non-specific binding of lactadherin may result in markedly increased counts. For all three labels, dual labeling for phosphatidylserine and leukocyte-, platelet-, or erythrocyte-specific markers is recommended to most accurately discriminate microparticle and non-microparticle events. The higher microparticle counts in erythrocyte concentrates compared to supernatants and more linear rate of development may more accurately reflect microparticle accumulations in erythrocyte concentrates for transfusion, but further studies are warranted to fully evaluate the role of non-specific binding in protein-rich and cell-rich erythrocyte concentrates.
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Details
- Title
- Comparison of established and novel methods for the detection and enumeration of microparticles in canine stored erythrocyte concentrates for transfusion
- Creators
- Meredeth Crandall McEntire
- Contributors
- K. Jane Wardrop (Degree Supervisor)
- Awarding Institution
- Washington State University
- Academic Unit
- Veterinary Medicine, College of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University; [Pullman, Washington] :
- Identifiers
- 99900525193301842
- Language
- English
- Resource Type
- Thesis