Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentrates
Abstract The extracellular vesicle release in red blood cell concentrates reflects progressive accumulation of storage lesions and could represent a new measure to be implemented routinely in blood centres in addition to haemolysis. Nevertheless, there is currently no standardized isolation protocol...
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| Format: | Article |
| Language: | English |
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Wiley
2025-01-01
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| Series: | Journal of Extracellular Biology |
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| Online Access: | https://doi.org/10.1002/jex2.70028 |
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| author | Marine Ghodsi Anne‐Sophie Cloos Anaïs Lotens Marine De Bueger Patrick Van Der Smissen Patrick Henriet Nicolas Cellier Christophe E. Pierreux Tomé Najdovski Donatienne Tyteca |
| author_facet | Marine Ghodsi Anne‐Sophie Cloos Anaïs Lotens Marine De Bueger Patrick Van Der Smissen Patrick Henriet Nicolas Cellier Christophe E. Pierreux Tomé Najdovski Donatienne Tyteca |
| author_sort | Marine Ghodsi |
| collection | DOAJ |
| description | Abstract The extracellular vesicle release in red blood cell concentrates reflects progressive accumulation of storage lesions and could represent a new measure to be implemented routinely in blood centres in addition to haemolysis. Nevertheless, there is currently no standardized isolation protocol. In a previous publication, we developed a reproducible ultracentrifugation‐based protocol (20,000 × g protocol) that allows to classify red blood cell concentrates into three cohorts according to their vesiculation level. Since this protocol was not adapted to meet routine requirements, the goal of this study was to develop an easier method based on low‐speed centrifugation (2,000 × g protocol) and limited red blood cell concentrate volumes to match with a non‐destructive sampling from the quality control sampling tubing. Despite the presence of contaminants, mainly in the form of albumin and lipoproteins, the material isolated with the 2,000 × g protocol contained red blood cell‐derived vesicular structures. It was reproducible, could predict the number of extracellular vesicles obtained with the 20,000 × g protocol and better discriminated between the three vesiculation cohorts than haemolysis at the legal expiry date of 6 weeks. However, by decreasing red blood cell concentrate volumes to fit with the volume in the quality control tubing, particle yield was highly reduced. Therefore, centrifugation time and relative centrifugal force were adapted (1,000 × g protocol), allowing for the recovery of a similar particle number and composition between small and large volumes sampled from the main unit, in different vesiculation cohorts over time. A similar observation was made with the 1,000 × g protocol between small volumes sampled from the quality control tubing and the mother‐bag. In conclusion, our study paves the way for the use of the 2,000 × g protocol (adapted to a 1,000 × g protocol with the quality control sampling tubing) for particle measurement in blood centres. |
| format | Article |
| id | doaj-art-e9b89a62bbb94e6fb3e773c16a4df9ff |
| institution | Kabale University |
| issn | 2768-2811 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Biology |
| spelling | doaj-art-e9b89a62bbb94e6fb3e773c16a4df9ff2025-01-27T13:48:44ZengWileyJournal of Extracellular Biology2768-28112025-01-0141n/an/a10.1002/jex2.70028Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentratesMarine Ghodsi0Anne‐Sophie Cloos1Anaïs Lotens2Marine De Bueger3Patrick Van Der Smissen4Patrick Henriet5Nicolas Cellier6Christophe E. Pierreux7Tomé Najdovski8Donatienne Tyteca9Cell Biology Unit & Platform for Imaging Cells and Tissues, de Duve Institute UCLouvain Brussels BelgiumCell Biology Unit & Platform for Imaging Cells and Tissues, de Duve Institute UCLouvain Brussels BelgiumService du Sang Croix‐Rouge de Belgique Suarlée BelgiumCell Biology Unit & Platform for Imaging Cells and Tissues, de Duve Institute UCLouvain Brussels BelgiumCell Biology Unit & Platform for Imaging Cells and Tissues, de Duve Institute UCLouvain Brussels BelgiumCell Biology Unit & Platform for Imaging Cells and Tissues, de Duve Institute UCLouvain Brussels BelgiumService du Sang Croix‐Rouge de Belgique Suarlée BelgiumCell Biology Unit & Platform for Imaging Cells and Tissues, de Duve Institute UCLouvain Brussels BelgiumService du Sang Croix‐Rouge de Belgique Suarlée BelgiumCell Biology Unit & Platform for Imaging Cells and Tissues, de Duve Institute UCLouvain Brussels BelgiumAbstract The extracellular vesicle release in red blood cell concentrates reflects progressive accumulation of storage lesions and could represent a new measure to be implemented routinely in blood centres in addition to haemolysis. Nevertheless, there is currently no standardized isolation protocol. In a previous publication, we developed a reproducible ultracentrifugation‐based protocol (20,000 × g protocol) that allows to classify red blood cell concentrates into three cohorts according to their vesiculation level. Since this protocol was not adapted to meet routine requirements, the goal of this study was to develop an easier method based on low‐speed centrifugation (2,000 × g protocol) and limited red blood cell concentrate volumes to match with a non‐destructive sampling from the quality control sampling tubing. Despite the presence of contaminants, mainly in the form of albumin and lipoproteins, the material isolated with the 2,000 × g protocol contained red blood cell‐derived vesicular structures. It was reproducible, could predict the number of extracellular vesicles obtained with the 20,000 × g protocol and better discriminated between the three vesiculation cohorts than haemolysis at the legal expiry date of 6 weeks. However, by decreasing red blood cell concentrate volumes to fit with the volume in the quality control tubing, particle yield was highly reduced. Therefore, centrifugation time and relative centrifugal force were adapted (1,000 × g protocol), allowing for the recovery of a similar particle number and composition between small and large volumes sampled from the main unit, in different vesiculation cohorts over time. A similar observation was made with the 1,000 × g protocol between small volumes sampled from the quality control tubing and the mother‐bag. In conclusion, our study paves the way for the use of the 2,000 × g protocol (adapted to a 1,000 × g protocol with the quality control sampling tubing) for particle measurement in blood centres.https://doi.org/10.1002/jex2.70028blood centresCoomassie blue gel stainingextracellular particlesextracellular vesiclesnanoparticle tracking analysis(ultra)centrifugation |
| spellingShingle | Marine Ghodsi Anne‐Sophie Cloos Anaïs Lotens Marine De Bueger Patrick Van Der Smissen Patrick Henriet Nicolas Cellier Christophe E. Pierreux Tomé Najdovski Donatienne Tyteca Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentrates Journal of Extracellular Biology blood centres Coomassie blue gel staining extracellular particles extracellular vesicles nanoparticle tracking analysis (ultra)centrifugation |
| title | Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentrates |
| title_full | Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentrates |
| title_fullStr | Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentrates |
| title_full_unstemmed | Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentrates |
| title_short | Development of an easy non‐destructive particle isolation protocol for quality control of red blood cell concentrates |
| title_sort | development of an easy non destructive particle isolation protocol for quality control of red blood cell concentrates |
| topic | blood centres Coomassie blue gel staining extracellular particles extracellular vesicles nanoparticle tracking analysis (ultra)centrifugation |
| url | https://doi.org/10.1002/jex2.70028 |
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