Extracellular vesicles from II trimester human amniotic fluid as paracrine conveyors counteracting oxidative stress.

Amniotic fluid Cell viability Extracellular vesicles Metabolic dysfunction Oxidative stress Paracrine effect

Journal

Redox biology
ISSN: 2213-2317
Titre abrégé: Redox Biol
Pays: Netherlands
ID NLM: 101605639

Informations de publication

Date de publication:
17 Jun 2024
Historique:
received: 20 04 2024
revised: 07 06 2024
accepted: 15 06 2024
medline: 21 6 2024
pubmed: 21 6 2024
entrez: 20 6 2024
Statut: aheadofprint

Résumé

We previously demonstrated that the human amniotic fluid (hAF) from II trimester of gestation is a feasible source of stromal progenitors (human amniotic fluid stem cells, hAFSC), with significant paracrine potential for regenerative medicine. Extracellular vesicles (EVs) separated and concentrated from hAFSC secretome can deliver pro-survival, proliferative, anti-fibrotic and cardioprotective effects in preclinical models of skeletal and cardiac muscle injury. While hAFSC-EVs isolation can be significantly influenced by in vitro cell culture, here we profiled EVs directly concentrated from hAF as an alternative option and investigated their paracrine potential against oxidative stress. II trimester hAF samples were obtained as leftover material from prenatal diagnostic amniocentesis following written informed consent. EVs were separated by size exclusion chromatography and concentrated by ultracentrifugation. hAF-EVs were assessed by nanoparticle tracking analysis, transmission electron microscopy, Western Blot, and flow cytometry; their metabolic activity was evaluated by oximetric and luminometric analyses and their cargo profiled by proteomics and RNA sequencing. hAF-EV paracrine potential was tested in preclinical in vitro models of oxidative stress and dysfunction on murine C2C12 cells and on 3D human cardiac microtissue. Our protocol resulted in a yield of 6.31 ± 0.98 × 10 Our results suggest that leftover samples of II trimester human amniotic fluid can represent a feasible source of EVs to counteract oxidative damage on target cells, thus offering a novel candidate therapeutic option to counteract skeletal and cardiac muscle injury.

Sections du résumé

BACKGROUND BACKGROUND
We previously demonstrated that the human amniotic fluid (hAF) from II trimester of gestation is a feasible source of stromal progenitors (human amniotic fluid stem cells, hAFSC), with significant paracrine potential for regenerative medicine. Extracellular vesicles (EVs) separated and concentrated from hAFSC secretome can deliver pro-survival, proliferative, anti-fibrotic and cardioprotective effects in preclinical models of skeletal and cardiac muscle injury. While hAFSC-EVs isolation can be significantly influenced by in vitro cell culture, here we profiled EVs directly concentrated from hAF as an alternative option and investigated their paracrine potential against oxidative stress.
METHODS METHODS
II trimester hAF samples were obtained as leftover material from prenatal diagnostic amniocentesis following written informed consent. EVs were separated by size exclusion chromatography and concentrated by ultracentrifugation. hAF-EVs were assessed by nanoparticle tracking analysis, transmission electron microscopy, Western Blot, and flow cytometry; their metabolic activity was evaluated by oximetric and luminometric analyses and their cargo profiled by proteomics and RNA sequencing. hAF-EV paracrine potential was tested in preclinical in vitro models of oxidative stress and dysfunction on murine C2C12 cells and on 3D human cardiac microtissue.
RESULTS RESULTS
Our protocol resulted in a yield of 6.31 ± 0.98 × 10
CONCLUSIONS CONCLUSIONS
Our results suggest that leftover samples of II trimester human amniotic fluid can represent a feasible source of EVs to counteract oxidative damage on target cells, thus offering a novel candidate therapeutic option to counteract skeletal and cardiac muscle injury.

Identifiants

DOI: 10.1016/j.redox.2024.103241 PMID: 38901103
pubmed: 38901103
pii: S2213-2317(24)00219-2
doi: 10.1016/j.redox.2024.103241
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

103241

Informations de copyright

Copyright © 2024. Published by Elsevier B.V.

Déclaration de conflit d'intérêts

Declaration of competing interest The authors have nothing to disclose nor competing interests to declare.

Auteurs

Senesi Giorgia (S)

Cardiovascular Theranostics, Istituto Cardiocentro Ticino and Laboratories for Traslational Research Ente Ospedaliero Cantonale, CH-6500, Bellinzona, Switzerland; Euler Institute, Faculty of Biomedical Sciences, Università della Svizzera Italiana, CH-6900, Lugano, Switzerland.

Guerricchio Laura (G)

Department of Experimental Medicine (DIMES), University of Genova, 16132, Genova, Italy.

Ghelardoni Maddalena (G)

IRCCS Ospedale Policlinico San Martino, 16132, Genova, Italy.

Bertola Nadia (B)

IRCCS Ospedale Policlinico San Martino, 16132, Genova, Italy.

Rebellato Stefano (R)

Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, 20900, Monza, Italy; School of Medicine and Surgery, University of Milano-Bicocca, 20900, Monza, Italy.

Grinovero Nicole (G)

Core Facilities - Clinical Proteomics and Metabolomics, IRCCS Istituto Giannina Gaslini, 16147, Genova, Italy.

Bartolucci Martina (B)

Core Facilities - Clinical Proteomics and Metabolomics, IRCCS Istituto Giannina Gaslini, 16147, Genova, Italy.

Costa Ambra (C)

IRCCS Ospedale Policlinico San Martino, 16132, Genova, Italy.

Raimondi Andrea (R)

Institute for Research in Biomedicine, Università della Svizzera Italiana, CH-6500, Bellinzona, Switzerland.

Grange Cristina (G)

VEXTRA Facility and Department of Medical Sciences, University of Turin, 10126, Turin, Italy.

Bolis Sara (B)

Cardiovascular Theranostics, Istituto Cardiocentro Ticino and Laboratories for Traslational Research Ente Ospedaliero Cantonale, CH-6500, Bellinzona, Switzerland.

Massa Valentina (M)

Department of Health Sciences, University of Milan, 20146, Milan, Italy.

Paladini Dario (P)

Fetal Medicine and Surgery Unit, IRCCS Istituto Giannina Gaslini, 16147, Genova, Italy.

Coviello Domenico (C)

Human Genetics Laboratory, IRCCS Istituto Giannina Gaslini, 16147, Genova, Italy.

Pandolfi Assunta (P)

Department of Medical, Oral and Biotechnological Sciences, University "G. d'Annunzio" Chieti-Pescara and Center for Advanced Studies and Technology - CAST, 66100, Chieti, Italy.

Bussolati Benedetta (B)

Department of Molecular Biotechnology and Health Sciences, University of Turin, 10126, Turin, Italy.

Petretto Andrea (P)

Core Facilities - Clinical Proteomics and Metabolomics, IRCCS Istituto Giannina Gaslini, 16147, Genova, Italy.

Fazio Grazia (F)

Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, 20900, Monza, Italy; School of Medicine and Surgery, University of Milano-Bicocca, 20900, Monza, Italy.

Ravera Silvia (R)

Department of Experimental Medicine (DIMES), University of Genova, 16132, Genova, Italy.

Barile Lucio (B)

Cardiovascular Theranostics, Istituto Cardiocentro Ticino and Laboratories for Traslational Research Ente Ospedaliero Cantonale, CH-6500, Bellinzona, Switzerland; Euler Institute, Faculty of Biomedical Sciences, Università della Svizzera Italiana, CH-6900, Lugano, Switzerland. Electronic address: lucio.barile@eoc.ch.

Balbi Carolina (B)

Center for Molecular Cardiology, University of Zurich, 8952, Schlieren, Switzerland; Department of Internal Medicine, Cantonal Hospital Baden, Baden, Switzerland. Electronic address: carolina.balbi@uzh.ch.

Bollini Sveva (B)

Department of Experimental Medicine (DIMES), University of Genova, 16132, Genova, Italy; IRCCS Ospedale Policlinico San Martino, 16132, Genova, Italy. Electronic address: sveva.bollini@unige.it.

Classifications MeSH