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https://doi.org/10.1186/s12964-024-01485-1
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Título: | Immunophenotype profile by flow cytometry reveals different subtypes of extracellular vesicles in porcine seminal plasma |
Fecha de publicación: | 23-ene-2024 |
Cita bibliográfica: | Cell Communication and Signaling. 2024. 22(1):63. |
ISSN: | Electronic: 1478-811X |
Palabras clave: | Seminal plasma Pig Extracellular vesicles Semen Reproduction Flow cytometry Ejaculate |
Resumen: | Background: Porcine seminal plasma (SP) is endowed with a heterogeneous population of extracellular vesicles (sEVs). This study evaluated the immunophenotypic profile by high-sensitivity flow cytometry of eight sEV subpopulations isolated according to their size (small [S-sEVs] and large [L-sEVs]) from four different SP sources, namely three ejaculate fractions (the first 10 mL of the sperm rich fraction [SRF-P1], the remaining SRF [SRF-P2], and the post-SRF [PSRF]) and entire ejaculate (EE). Methods: Seminal EVs were isolated using a size exclusion chromatography-based protocol from six SP pools (five ejaculates/pool) of each SP source and characterized using complementary approaches including total protein (BCA™assay), particle size distribution (dynamic light scattering), morphology (transmission electron microscopy), and purity (albumin by Western blot). Expression of CD9, CD63, CD81, CD44 and HSP90β was analyzed in all sEV subpopulations by high-sensitivity flow cytometry according to MIFlowCyt-EV guidelines, including an accurate calibration, controls, and discrimination by CFSE-labelling. Results: Each sEV subpopulation exhibited a specific immunophenotypic profile. The percentage of sEVs positive for CD9, CD63, CD81 and HSP90β differed between S- and L-sEVs (P < 0.0001). Specifically, the percentage of sEVs positive for CD9 and CD63 was higher and that for CD81 was lower in S- than L-sEVs in the four SP sources. However, the percentage of HSP90β-positive sEVs was lower in S-sEVs than L-sEVs in the SRF-P1 and EE samples. The percentage of sEVs positive for CD9, CD63, and CD44 also differed among the four SP sources (P < 0.0001), being highest in PSRF samples. Notably, virtually all sEV subpopulations expressed CD44 (range: 88.04-98.50%). Conclusions: This study demonstrated the utility of high-sensitivity flow cytometry for sEV immunophenotyping, allowing the identification of distinct sEV subpopulations that may have different cellular origin, cargo, functions, and target cells. |
Autor/es principal/es: | Barranco, Isabel Alvarez-Barrientos, Alberto Parra, Ana Martinez-Diaz, Pablo Lucas, Xiomara Roca, Jordi |
Facultad/Departamentos/Servicios: | Facultades, Departamentos, Servicios y Escuelas::Departamentos de la UMU::Medicina y Cirugía Animal Facultades, Departamentos, Servicios y Escuelas::Facultades de la UMU::Facultad de Veterinaria |
Versión del editor: | https://biosignaling.biomedcentral.com/articles/10.1186/s12964-024-01485-1#Fun |
URI: | http://hdl.handle.net/10201/140143 |
DOI: | https://doi.org/10.1186/s12964-024-01485-1 |
Tipo de documento: | info:eu-repo/semantics/article |
Número páginas / Extensión: | 17 |
Derechos: | info:eu-repo/semantics/openAccess Atribución 4.0 Internacional |
Descripción: | © 2024. The authors. This document is made available under the CC-BY 4.0 license http://creativecommons.org/licenses/by /4.0/ This document is the published version of a published work that appeared in final form in Cell Communication and Signaling (CCS). To access the final work, see DOI: https://doi.org/10.1186/s12964-024-01485-1 |
Aparece en las colecciones: | Artículos: Medicina y Cirugía Animal |
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