Isolation of endothelial colony-forming cells from blood samples collected from the jugular and cephalic veins of healthy adult horses.
Abstract: OBJECTIVE To evaluate optimal isolation of endothelial colony-forming cells (ECFCs) from peripheral blood of horses. SAMPLE Jugular and cephalic venous blood samples from 17 adult horses. PROCEDURES Each blood sample was divided; isolation was performed with whole blood adherence (WBA) and density gradient centrifugation (DGC). Isolated cells were characterized by uptake of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate-labeled acetylated low-density lipoprotein (DiI-Ac-LDL), vascular tubule formation, and expression of endothelial (CD34, CD105, vascular endothelial growth factor receptor-2, and von Willebrand factor) and hematopoietic (CD14) cell markers by use of indirect immunofluorescence assay (IFA) and flow cytometry. RESULTS Colonies with cobblestone morphology were isolated from 15 of 17 horses. Blood collected from the cephalic vein yielded colonies significantly more often (14/17 horses) than did blood collected from the jugular vein (8/17 horses). Of 14 cephalic blood samples with colonies, 13 were obtained with DGC and 8 with WBA. Of 8 jugular blood samples with colonies, 8 were obtained with DGC and 4 with WBA. Colony frequency (colonies per milliliter of blood) was significantly higher for cephalic blood samples and samples isolated with DGC. Cells formed vascular tubules, had uptake of DiI-Ac-LDL, and expressed endothelial markers by use of IFA and flow cytometry, which confirmed their identity as ECFCs. CONCLUSIONS AND CLINICAL RELEVANCE Maximum yield of ECFCs was obtained for blood samples collected from both the jugular and cephalic veins and use of DGC to isolate cells. Consistent yield of ECFCs from peripheral blood of horses will enable studies to evaluate diagnostic and therapeutic uses.
Publication Date: 2016-09-27 PubMed ID: 27668588DOI: 10.2460/ajvr.77.10.1157Google Scholar: Lookup
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- Journal Article
Summary
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The research article is about the successful isolation of endothelial colony-forming cells (ECFCs) from horse blood drawn from jugular and cephalic veins, with an increase in yield when using density gradient centrifugation.
Introduction
- The research aims to establish an efficient method of isolating endothelial colony-forming cells (ECFCs) from the blood of adult horses.
- These ECFCs are important in the construction and repair of blood vessels. Understanding how to effectively isolate them can contribute to veterinary and potentially human medical advancement.
Methodology
- The blood samples were obtained from the jugular and cephalic veins of 17 healthy adult horses.
- The blood samples were divided and the isolation process was performed through two methods: whole blood adherence (WBA) and density gradient centrifugation (DGC).
- The isolated cells were characterized based on the uptake of DiI-Ac-LDL, vascular tubule formation, and expression of endothelial and hematopoietic cell markers. The expressed markers were evaluated through indirect immunofluorescence assay (IFA) and flow cytometry.
Results
- Colonies with cobblestone morphology were identified in samples from 15 of the 17 horses.
- The yield from cephalic blood samples was significantly higher (14 out of 17 horses) compared to that from jugular samples (8 out of 17 horses).
- The number of colonies per milliliter of blood (colony frequency) was notably higher for the cephalic blood and samples isolated through DGC.
- Results confirmed the isolated cells as ECFCs, hence the method used was deemed successful.
Conclusion and Clinical Relevance
- The most successful isolation of ECFCs occurred with blood samples from both the jugular and cephalic veins using DGC.
- Consistent yield of ECFCs from peripheral blood samples of horses could enable further studies for diagnostic and potential therapeutic uses.
Cite This Article
APA
Sharpe AN, Seeto WJ, Winter RL, Zhong Q, Lipke EA, Wooldridge AA.
(2016).
Isolation of endothelial colony-forming cells from blood samples collected from the jugular and cephalic veins of healthy adult horses.
Am J Vet Res, 77(10), 1157-1165.
https://doi.org/10.2460/ajvr.77.10.1157 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Brachiocephalic Veins / diagnostic imaging
- Endothelial Cells / cytology
- Female
- Flow Cytometry / veterinary
- Fluorescent Antibody Technique / veterinary
- Horses
- Jugular Veins / diagnostic imaging
- Male
- Reference Values
- Vascular Endothelial Growth Factor A / metabolism
- von Willebrand Factor / metabolism
Citations
This article has been cited 3 times.- Winter RL, Tian Y, Caldwell FJ, Seeto WJ, Koehler JW, Pascoe DA, Fan S, Gaillard P, Lipke EA, Wooldridge AA. Cell engraftment, vascularization, and inflammation after treatment of equine distal limb wounds with endothelial colony forming cells encapsulated within hydrogel microspheres. BMC Vet Res 2020 Feb 4;16(1):43.
- Winter RL, Seeto WJ, Tian Y, Caldwell FJ, Lipke EA, Wooldridge AA. Growth and function of equine endothelial colony forming cells labeled with semiconductor quantum dots. BMC Vet Res 2018 Aug 23;14(1):247.
- Finding EJT, Faulkner A, Nash L, Wheeler-Jones CPD. Equine Endothelial Cells Show Pro-Angiogenic Behaviours in Response to Fibroblast Growth Factor 2 but Not Vascular Endothelial Growth Factor A. Int J Mol Sci 2024 May 30;25(11).
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