Extracellular Vesicles Core Facility (UVEx)

Extracellular Vesicles Core Facility (UVEx)

 

 

Core director: Virginia Vila del Sol

E-mail: UVEx.hnp@sescam.jccm.es

 

 

Extracellular Vesicles (EVs) are a heterogeneous population of small-sized membrane-round vesicleswhich are secreted by cells in physiological and/or pathological conditions. EVs show a wide variety of functions and biological properties mainly depending on their cargo, composed by bioactive molecules such as proteins, lipids and nucleic acids, especially non-coding RNA (miRNA, IncRNA, CircRNA). EVs might be found on almost all the biofluids: plasma, urine, saliva, breast milk, amniotic fluid, cerebrospinal fluid, seminal plasma, etc. In addition to their role as messengers on intercellular communication by transporting information and material from cell to cell, EVs have an importantrole on cellular migration processes, angiogenesis andtumor growing.

 

Attending to all these special characteristics, the analysis of EVs as biomarkers as well as drug-delivery vehicles, is gaining importance in the last years, although their characterization needs forstate-of-the-art techniques and a highly qualified staff specialized in diverse technologies, who work by following two main guidelines: MISEV 2023 and MIFlowCyt, which are recommendations for the standardization and reporting of EVs studies.

 

In 2022, the Flow Cytometry and the Proteomics core facilities were associated in a new Interdisciplinary Core Facility specialized in the study of EVs to provide researchers the necessary technologies from the first step of EVs enrichment to the last one for protein cargo characterization.

 

Recently, UVEx acquired a new platform for exosome quantification and characterization: the ExoView R100 system. This system enables the direct analysis of EVs in biofluids (such as plasma, urine, cerebrospinal fluid, among others) without the need of prior isolation or purification steps. The ExoView R100 technology opens up new possibilities for bringing biomedical research closer to clinical application, allowing for disease monitoring and the development of new therapies.

 

Moreover, since 2021, UVEx belongs to “Grupo Español de Innovación e Investigación en Vesículas Extracelulares”-- GEIVEX. GEIVEX is a Spanish organization aimed to promote the research, innovation and knowledge exchange between research groups in the field of EVs.

 

 

 

 

Technical assays

 

All the offered assays are performed by following MISEV 2023 and MIFlowCyt-EVs guidelines

 

 

Isolation and enrichment of Extracellular Vesicles

- Differential ultracentrifugation (dUC).  
- Immunocapture (IC).
- Size Exclusion Chromatography (SEC).
- Ultrafiltration (UF).

EV characterization by ExoView R100

- EV quantification (particles/ml)
- Single nanoparticle size determination
- Immunophenotype characterization and biomarker colocalization analysis
- Simultaneous EVs cargo determination
- Lentiviral titration
- Up to 35 µl sample volume without prior purification

 

EVs Characterization by Flow Cytometry

- EV Quantification (events/µl).
- Immunocapture (IC). 
- Direct detection of EVs-specific markers: CD63, CD81 and CD9 tetraspanins.
- Direct detection of cell-of-origin-specific markers in EVs.

 

 

 

Characterization of EVs by WB

- Characterization of transmembrane proteins by Western Blot (WB): CD9, CD81 y CD63.  
- Characterization of cytoplasmic proteins by Western Blot (WB): HSP70, TSG101 ALIX.

 

 

 

 

Extracellular vesicles analysis by MS

- Identification of EVs protein profile by mass spectrometry (MS). 
- Differential expression analysis of EVsproteins.
- Extracellular vesicles metabolomics analysis.
- Extracellular vesicles lipidomic analysis.

 

 

 

 

 

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Publications and recent communications

 

- Vila del Sol, Virginia; Marquina Rodríguez, Ángela; González Arandilla, Alba ; Mendoza De Mingo, Patricia ; González López, Mª José ; Gallardo Escobar, Ana; Barroso García, Gemma “Unidad de vesículas extracelulares: herramienta científico-técnica para el estudio de vesículas extracelulares en medicina personalizada de precisión” Póster Jornada de Investigación de IDISCAM, 02 diciembre 2024

 

- Mª Cristina Ortega, Cristina Martos-Polo, Pilar García-Llorena, Rocío del C. Bravo-Miana, Ángela Marquina-Rodríguez, José Alberto Suárez, Virginia Vila-del Sol, David Otaegui, Diego Clemente “Characterization of extracellular vesicles from myeloid-derived suppressor cells: role in the severity of experimental multiple sclerosis” Póster 7th GEIVEX Symposium. October 23-25, 2024.

 

-Mª Cristina Ortega, Pilar García-Llorena, Cristina Martos-Polo, José Alberto Suárez, Jénnifer García-Arocha, Ángela Marquina-Rodríguez, Virginia Vila-del Sol, David Otaegui y Diego Clemente “Caracterización de las vesículas extracelulares procedentes de las células mieloides supresoras: relación con la severidad clínica en la esclerosis múltiple experimental” Póster II Jornadas de Investigación Sanitaria IDISCAM. 14 Febrero 2024.

 

- Marquina-Rodríguez A., Ortega MC, García-Llorena P., Barroso-García G. Arandilla AG, González MJ, Clemente D, Vila-del Sol V. “Extracellular Vesicles from sorted Myeloid-Derived Suppressor Cells mantain specific staining from sorting". Poster exhibition at XVIII Congress of the IBERIAN SOCIETY of CYTOMETRY. April 19-21, 2023 (Abstract SIC 2023).

 

- Marquina-Rodríguez A., Barroso-García G., Arandilla AG, Ortega MC, García-Llorena P, Clemente D, Vila-del Sol V. “Implementation of an Interdisciplinary Core Facility for the study of Extracellular Vesicles at the National Hospital of Paraplegics”. Poster exhibition at 6th GEIVEX Symposium. October 25-28, 2022 (Abstract 6th Symposium GEIVEX).

 

-Serrano-Heras, G., Díaz-Maroto, I., Castro-Robles, B. et al. “Isolation and Quantification of Blood Apoptotic Bodies, a Non-invasive Tool to Evaluate Apoptosis in Patients with Ischemic Stroke and Neurodegenerative Diseases”. BiolProced Online 22, 17 (2020).
https://doi.org/10.1186/s12575-020-00130-8.

 

 

 

Staff

 

Core Director:

 

- Virginia Vila del Sol, PhD
Email: vvila@sescam.jccm.es

 

 

Research Technicians:

 

- Ángela Marquina Rodríguez, MSc.
Email: amarquinar@sescam.jccm.es

 

- Gemma Barroso García, MSc.
Email: gbarroso@sescam.jccm.es

 

- Alba González Arandilla
Email: agarandilla@sescam.jccm.es

 

- Inmaculada Fernández-Cañadas Sánchez-Aguilera 
Email: ifernandezcanadas@sescam.jccm.es

 

 

Lab Technicians:

 

- María José González López.
Email: mjgonzalezl@sescam.jccm.es

 

- Ana Gallardo Escobar.
Email: agallardoe@sescam.jccm.es

 

 

Contact

 

Extracellular Vesicles core facility-UVEx
Edif. Investigación i-1. Lab. 23.
Hospital Nacional de Parapléjicos
Finca La Peraleda s/n
45071-Toledo
Spain
Email: UVEx.hnp@sescam.jccm.es
Phone. 1: (+34) 925 396 833
Phone. 2: (+34) 925 396 823
Fax: (+34) 925 396 821