Methodological Assessment of ExoGAG for Isolation of Cerebrospinal Fluid Extracellular Vesicles as a Source of Biomarkers
Por:
Salvat-Rovira, N, Vazquez-Oliver, A, Rivas-Asensio, E, Herrero-Lorenzo, M, Gámez-Valero, A, Pérez-Pérez, J, Izquierdo, C, Campolongo, A, Marti, E, Kulisevsky, J, Pérez-González, R
Publicada:
1 dic 2024
Resumen:
Extracellular vesicles (EVs) in cerebrospinal fluid (CSF) represent a valuable source of biomarkers for central nervous system (CNS) diseases, offering new pathways for diagnosis and monitoring. However, existing methods for isolating EVs from CSF often prove to be labor-intensive and reliant on specialized equipment, hindering their clinical application. In this study, we present a novel, clinically compatible method for isolating EVs from CSF. We optimized the use of ExoGAG, a commercially available reagent that has been tested in plasma, urine and semen, and compared it directly with differential ultracentrifugation using Western blotting, protein quantification, nanoparticle tracking analysis, and cryogenic electron microscopy. Additionally, we analyzed the presence of specific microRNAs (miRNAs) known to be present in CSF-derived EVs. Our data demonstrate that ExoGAG is an effective method for isolating EVs from CSF, yielding a higher amount of EVs compared to traditional ultracentrifugation methods, and with comparable levels of specific miRNAs. In conclusion, the use of ExoGAG in a clinical setting may facilitate the testing of biomarkers essential for tracking brain pathology in CNS diseases.
Filiaciones:
Salvat-Rovira, N:
Hosp Santa Creu i Sant Pau, Neurol Dept, Movement Disorders Unit, Barcelona 08025, Spain
Sant Pau Biomed Res Inst IIB Sant Pau, Barcelona 08041, Spain
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Univ Autonoma Barcelona, Dept Med, Bellaterra 08193, Spain
Vazquez-Oliver, A:
Hosp Santa Creu i Sant Pau, Neurol Dept, Movement Disorders Unit, Barcelona 08025, Spain
Sant Pau Biomed Res Inst IIB Sant Pau, Barcelona 08041, Spain
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Rivas-Asensio, E:
Hosp Santa Creu i Sant Pau, Neurol Dept, Movement Disorders Unit, Barcelona 08025, Spain
Sant Pau Biomed Res Inst IIB Sant Pau, Barcelona 08041, Spain
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Herrero-Lorenzo, M:
Univ Barcelona, Inst Neurosci, Fac Med, Dept Biomed, Barcelona 08036, Spain
Gámez-Valero, A:
Univ Barcelona, Inst Neurosci, Fac Med, Dept Biomed, Barcelona 08036, Spain
Pérez-Pérez, J:
Hosp Santa Creu i Sant Pau, Neurol Dept, Movement Disorders Unit, Barcelona 08025, Spain
Sant Pau Biomed Res Inst IIB Sant Pau, Barcelona 08041, Spain
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Izquierdo, C:
Hosp Santa Creu i Sant Pau, Neurol Dept, Movement Disorders Unit, Barcelona 08025, Spain
Sant Pau Biomed Res Inst IIB Sant Pau, Barcelona 08041, Spain
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Campolongo, A:
Hosp Santa Creu i Sant Pau, Neurol Dept, Movement Disorders Unit, Barcelona 08025, Spain
Sant Pau Biomed Res Inst IIB Sant Pau, Barcelona 08041, Spain
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Marti, E:
Univ Barcelona, Inst Neurosci, Fac Med, Dept Biomed, Barcelona 08036, Spain
Kulisevsky, J:
Hosp Santa Creu i Sant Pau, Neurol Dept, Movement Disorders Unit, Barcelona 08025, Spain
Sant Pau Biomed Res Inst IIB Sant Pau, Barcelona 08041, Spain
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Pérez-González, R:
Ctr Invest Red Enfermedades Neurodegenerat CIBERNE, Madrid 19171, Spain
Inst Invest Sanitaria & Biomed Alicante ISABIAL, Alicante 03010, Spain
Univ Miguel Hernandez, CSIC, Inst Neurociencias, Sant Joan Dalacant 03550, Spain
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