Characterization and quantification of porcine circulating endothelial cells
| Autor: | Robert A. van Hulst, Albert C.W.A. van Wijk, Michal Heger, Robert P. Weenink, Ivo C.J.H. Post, Thomas M. van Gulik, Anita N. Böing |
|---|---|
| Přispěvatelé: | Other departments, Amsterdam Gastroenterology Endocrinology Metabolism, Cancer Center Amsterdam, 02 Surgical specialisms, Amsterdam Cardiovascular Sciences, Other Research, Laboratory for Experimental Clinical Chemistry, Surgery |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
CD31
Endothelium Immunology Sus scrofa Transplantation Heterologous Antibodies Heterophile Cell Count Receptors Cell Surface CD146 Antigen Biology Cross Reactions Epitope Umbilical vein Flow cytometry Antigens CD Antigens Heterophile medicine Human Umbilical Vein Endothelial Cells Animals Humans Cells Cultured Whole blood Transplantation Blood Cells Microscopy Confocal medicine.diagnostic_test Endoglin Endothelial Cells Flow Cytometry Molecular biology medicine.anatomical_structure cardiovascular system CD146 |
| Zdroj: | Xenotransplantation, 20(1), 18-26. Wiley-Blackwell |
| ISSN: | 0908-665X |
| Popis: | Background Endothelial damage is a critical step in the development of (xeno) transplantation-related and cardiovascular pathology. In humans, the amount of circulating endothelial cells (CEC) correlates to disease intensity and functions as a valuable damage marker. While (xeno) transplantation and cardiovascular research is regularly performed in porcine models, the paucity of antibodies against porcine endothelium epitopes hinders the use of CEC as damage marker. Objective This study aimed to develop a method for porcine CEC detection using anti-human antibodies against porcine endothelium epitopes. Methods Human umbilical vein endothelial cells (HUVEC, control) and their swine equivalent (SUVEC) were used to assess the cross-species immunoreactivity of fluorescently labeled anti-human CD31/CD51/CD54/CD62E/CD105/CD106/CD144/CD146/PAL-E/lectin-1/vWF antibodies by isotype-controlled fluorescence-activated cell sorting (FACS) and confocal microscopy. Next, reactivity was ascertained with mature porcine kidney-derived endothelial cells (PKEC), and a FACS-based whole blood CEC quantification method was employed using osmotic erythrolysis and CD105 and CD146 double staining after CD45 exclusion. Results Of the 21 assayed antibodies, the MEM-229 clone of CD105 and P1H12 clone of CD146 showed immunoreactivity with SUVEC and PKEC. Double staining showed baseline porcine CEC count of 673.1 ± 551.4 CEC/ml, while the first 7.5 ml of drawn blood (representative of vascular damage) contained 1118 ± 661.4 CEC/ml (n = 14, P = 0.04). A second experiment (n = 5) including CD45 exclusion identified only 14.5 ± 10.8% double-positive CD105-146 events per ml blood. Conclusion Porcine endothelium can be specifically labeled using anti-human CD146 and CD105 antibodies. These antibodies can therefore be used for the identification and quantification of CEC in porcine whole blood by FACS after osmotic erythrolysis. |
| Databáze: | OpenAIRE |
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