Immunofluorescence Protocol for Characterization of Platelet and Leukocyte Binding in Extracorporeal Membrane Oxygenation (ECMO) Circuits.

Autor:	Cai T; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia.; Department of Paediatrics, The University of Melbourne, Parkville, Australia., Burton M; Flow Cytometry and Imaging, Murdoch Children's Research Institute, Parkville, Australia., McCafferty C; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia.; Department of Paediatrics, The University of Melbourne, Parkville, Australia., Van Den Helm S; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia.; Department of Paediatrics, The University of Melbourne, Parkville, Australia., Letunica N; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia., Attard C; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia.; Department of Paediatrics, The University of Melbourne, Parkville, Australia., Horton S; Department of Paediatrics, The University of Melbourne, Parkville, Australia.; Cardiac Surgery, The Royal Children's Hospital, Parkville, Australia., Bottrell S; Cardiac Surgery, The Royal Children's Hospital, Parkville, Australia., Schultz B; Cardiac Surgery, The Royal Children's Hospital, Parkville, Australia., MacLaren G; Cardiothoracic Intensive Care Unit, National University Health System, Singapore., Chiletti R; Intensive Care, The Royal Children's Hospital, Parkville, Australia.; Paediatric Intensive Care Research Group, Murdoch Children's Research Institute, Parkville, Australia., Best D; Intensive Care, The Royal Children's Hospital, Parkville, Australia.; Paediatric Intensive Care Research Group, Murdoch Children's Research Institute, Parkville, Australia., Johansen A; Intensive Care, The Royal Children's Hospital, Parkville, Australia.; Paediatric Intensive Care Research Group, Murdoch Children's Research Institute, Parkville, Australia., Newall F; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia.; Department of Paediatrics, The University of Melbourne, Parkville, Australia.; Clinical Haematology, The Royal Children's Hospital, Parkville, Australia., Butt W; Department of Paediatrics, The University of Melbourne, Parkville, Australia.; Intensive Care, The Royal Children's Hospital, Parkville, Australia.; Paediatric Intensive Care Research Group, Murdoch Children's Research Institute, Parkville, Australia., d'Udekem Y; Cardiovascular Surgery, Children's National Heart Institute, Washington, DC., Monagle P; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia.; Department of Paediatrics, The University of Melbourne, Parkville, Australia.; Clinical Haematology, The Royal Children's Hospital, Parkville, Australia.; Kids Cancer Centre, Sydney Children's Hospital, Randwick, Australia., Ignjatovic V; From the Haematology Research, Murdoch Children's Research Institute, Parkville, Australia.; Department of Paediatrics, The University of Melbourne, Parkville, Australia.; Cardiac Surgery, The Royal Children's Hospital, Parkville, Australia.; Johns Hopkins All Children's Institute for Clinical and Translational Research, St Petersburg, Florida.; Department of Pediatrics, School of Medicine, Johns Hopkins University, Baltimore, Maryland.
Jazyk:	angličtina
Zdroj:	ASAIO journal (American Society for Artificial Internal Organs : 1992) [ASAIO J] 2025 Jan 01; Vol. 71 (1), pp. e1-e4. Date of Electronic Publication: 2024 Jun 21.
DOI:	10.1097/MAT.0000000000002259
Abstrakt:	The continuous contact between blood and the foreign surface of the extracorporeal membrane oxygenation (ECMO) circuit contributes to hemostatic, inflammatory, and other physiological disturbances observed during ECMO. Although previous studies have extensively investigated blood samples from patients on ECMO, cell adsorption to the ECMO circuit as an additional factor that could potentially influence clinical outcomes, has largely been overlooked. Here we provide a detailed immunofluorescence (IF) protocol designed to characterize cellular binding on ECMO circuits collected from patients. Extracorporeal membrane oxygenation circuits were collected from three pediatric patients and an albumin primed-only ECMO circuit was used as control. Circuit samples from five different sites within each ECMO circuit were collected and processed for the IF protocol. CD14 and CD42a antibodies were used to identify platelets and leukocytes bound to each ECMO circuit sample and images captured using inverted fluorescence microscopy. The protocol enables the comprehensive characterization of platelet and leukocyte binding to ECMO circuits collected from patients, which could in turn extend our knowledge of the characteristics of circuit binding and may provide guidance for improved ECMO circuit design. Competing Interests: G.M. is the president of the Extracorporeal Life Support Organization (ELSO). The other authors have no conflicts of interest to report. (Copyright © ASAIO 2024.)
Databáze:	MEDLINE
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