| Methodologic considerations for the use of canine in vivo aged biotinylated erythrocytes to study RBC senescence. | |
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MedLine Citation:
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PMID: 8536797 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Biotinylation of erythrocytes has been developed in rabbits as a tool to retrieve labeled cells following various periods in circulation. This retrieval capability allows biochemical studies to be conducted on red blood cells (RBC) that have aged for desired times in vivo. However, because erythrocyte life span is much shorter in rabbits than in humans, and because cell removal is measurably age-independent in rabbits, we have sought to validate the same protocol in dogs, whose cell life span and age-dependent removal characteristics are similar to humans'. Canine RBC were biotinylated in vivo by infusion of N-hydroxysuccinimidyl biotin dissolved in dimethylacetamide or dimethylsulfoxide. Cell life spans were evaluated using 14C-cyanate labeling followed by scintillation counting or avidin-FITC labeling followed by flow cytometry. Both methods gave identical results. The life span of the biotin-conjugated cells was found to be normal (approximately 110 days), and the stability of the biotin ligand was adequate for efficient retrieval of cells using avidin-coated magnetic beads (magnetic cell sorting [MACS]). From each isolation, approximately 20 microL of packed biotinylated cells of approximately 90% purity (i.e., 10% contamination by unlabeled cells) could be harvested. On average, approximately 60% of the biotinylated cells in any sample could be retrieved. Either multiple isolations or use of larger collection columns will facilitate collection of cell numbers sufficient for biochemical tests. After incorporating several modifications in the previous biotinylation protocol that were required for adaptation to the dog, the methodology can be used to study red cell senescence in an animal that has several pertinent similarities to humans. |
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Authors:
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J A Christian; A H Rebar; G D Boon; P S Low |
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Publication Detail:
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Type: Comparative Study; Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Experimental hematology Volume: 24 ISSN: 0301-472X ISO Abbreviation: Exp. Hematol. Publication Date: 1996 Jan |
Date Detail:
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Created Date: 1996-02-08 Completed Date: 1996-02-08 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 0402313 Medline TA: Exp Hematol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 82-8 Citation Subset: IM |
Affiliation:
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Department of Veterinary Pathobiology, Purdue University, West Lafayette, IN 47907-1243, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acetamides
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toxicity Animals Biotin / blood* Carbon Radioisotopes Dimethyl Sulfoxide Dogs Drug-Induced Liver Injury Erythrocyte Aging* Female Flow Cytometry Male Models, Biological Scintillation Counting Time Factors |
| Grant Support | |
ID/Acronym/Agency:
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GM24417/GM/NIGMS NIH HHS; HL08178/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Acetamides; 0/Carbon Radioisotopes; 127-19-5/dimethylacetamide; 58-85-5/Biotin; 67-68-5/Dimethyl Sulfoxide |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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