Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes

Alice Busato,1,* Roberta Bonafede,1,* Pietro Bontempi,2 Ilaria Scambi,1 Lorenzo Schiaffino,1 Donatella Benati,1 Manuela Malatesta,1 Andrea Sbarbati,1 Pasquina Marzola,3 Raffaella Mariotti1 1Department of Neurosciences, Biomedicine and Movement Sciences, School of Medicine, 2Department of Biotechnol...

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Autores principales: Busato A, Bonafede R, Bontempi P, Scambi I, Schiaffino L, Benati D, Malatesta M, Sbarbati A, Marzola P, Mariotti R
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Lenguaje:EN
Publicado: Dove Medical Press 2016
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MRI
Acceso en línea:https://doaj.org/article/88895595986b40bbb3bb87ba84d3d04c
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spelling oai:doaj.org-article:88895595986b40bbb3bb87ba84d3d04c2021-12-02T06:34:33ZMagnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes1178-2013https://doaj.org/article/88895595986b40bbb3bb87ba84d3d04c2016-06-01T00:00:00Zhttps://www.dovepress.com/magnetic-resonance-imaging-of-ultrasmall-superparamagnetic-iron-oxide--peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Alice Busato,1,* Roberta Bonafede,1,* Pietro Bontempi,2 Ilaria Scambi,1 Lorenzo Schiaffino,1 Donatella Benati,1 Manuela Malatesta,1 Andrea Sbarbati,1 Pasquina Marzola,3 Raffaella Mariotti1 1Department of Neurosciences, Biomedicine and Movement Sciences, School of Medicine, 2Department of Biotechnology, 3Department of Computer Science, University of Verona, Verona, Italy *These authors contributed equally to this work Purpose: Recent findings indicate that the beneficial effects of adipose stem cells (ASCs), reported in several neurodegenerative experimental models, could be due to their paracrine activity mediated by the release of exosomes. The aim of this study was the development and validation of an innovative exosome-labeling protocol that allows to visualize them with magnetic resonance imaging (MRI).Materials and methods: At first, ASCs were labeled using ultrasmall superparamagnetic iron oxide nanoparticles (USPIO, 4–6 nm), and optimal parameters to label ASCs in terms of cell viability, labeling efficiency, iron content, and magnetic resonance (MR) image contrast were investigated. Exosomes were then isolated from labeled ASCs using a standard isolation protocol. The efficiency of exosome labeling was assessed by acquiring MR images in vitro and in vivo as well as by determining their iron content. Transmission electron microscopy images and histological analysis were performed to validate the results obtained.Results: By using optimized experimental parameters for ASC labeling (200 µg Fe/mL of USPIO and 72 hours of incubation), it was possible to label 100% of the cells, while their viability remained comparable to unlabeled cells; the detection limit of MR images was of 102 and 2.5×103 ASCs in vitro and in vivo, respectively. Exosomes isolated from previously labeled ASCs retain nanoparticles, as demonstrated by transmission electron microscopy images. The detection limit by MRI was 3 µg and 5 µg of exosomes in vitro and in vivo, respectively.Conclusion: We report a new approach for labeling of exosomes by USPIO that allows detection by MRI while preserving their morphology and physiological characteristics. Keywords: MRI, superparamagnetic iron oxide nanoparticles, cellular imaging, stem cells labeling, exosome labelingBusato ABonafede RBontempi PScambi ISchiaffino LBenati DMalatesta MSbarbati AMarzola PMariotti RDove Medical PressarticleMRIsuperparamagnetic iron oxide nanoparticlescellular imagingstem cells labelingexosome labelingMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2016, Iss default, Pp 2481-2490 (2016)
institution DOAJ
collection DOAJ
language EN
topic MRI
superparamagnetic iron oxide nanoparticles
cellular imaging
stem cells labeling
exosome labeling
Medicine (General)
R5-920
spellingShingle MRI
superparamagnetic iron oxide nanoparticles
cellular imaging
stem cells labeling
exosome labeling
Medicine (General)
R5-920
Busato A
Bonafede R
Bontempi P
Scambi I
Schiaffino L
Benati D
Malatesta M
Sbarbati A
Marzola P
Mariotti R
Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes
description Alice Busato,1,* Roberta Bonafede,1,* Pietro Bontempi,2 Ilaria Scambi,1 Lorenzo Schiaffino,1 Donatella Benati,1 Manuela Malatesta,1 Andrea Sbarbati,1 Pasquina Marzola,3 Raffaella Mariotti1 1Department of Neurosciences, Biomedicine and Movement Sciences, School of Medicine, 2Department of Biotechnology, 3Department of Computer Science, University of Verona, Verona, Italy *These authors contributed equally to this work Purpose: Recent findings indicate that the beneficial effects of adipose stem cells (ASCs), reported in several neurodegenerative experimental models, could be due to their paracrine activity mediated by the release of exosomes. The aim of this study was the development and validation of an innovative exosome-labeling protocol that allows to visualize them with magnetic resonance imaging (MRI).Materials and methods: At first, ASCs were labeled using ultrasmall superparamagnetic iron oxide nanoparticles (USPIO, 4–6 nm), and optimal parameters to label ASCs in terms of cell viability, labeling efficiency, iron content, and magnetic resonance (MR) image contrast were investigated. Exosomes were then isolated from labeled ASCs using a standard isolation protocol. The efficiency of exosome labeling was assessed by acquiring MR images in vitro and in vivo as well as by determining their iron content. Transmission electron microscopy images and histological analysis were performed to validate the results obtained.Results: By using optimized experimental parameters for ASC labeling (200 µg Fe/mL of USPIO and 72 hours of incubation), it was possible to label 100% of the cells, while their viability remained comparable to unlabeled cells; the detection limit of MR images was of 102 and 2.5×103 ASCs in vitro and in vivo, respectively. Exosomes isolated from previously labeled ASCs retain nanoparticles, as demonstrated by transmission electron microscopy images. The detection limit by MRI was 3 µg and 5 µg of exosomes in vitro and in vivo, respectively.Conclusion: We report a new approach for labeling of exosomes by USPIO that allows detection by MRI while preserving their morphology and physiological characteristics. Keywords: MRI, superparamagnetic iron oxide nanoparticles, cellular imaging, stem cells labeling, exosome labeling
format article
author Busato A
Bonafede R
Bontempi P
Scambi I
Schiaffino L
Benati D
Malatesta M
Sbarbati A
Marzola P
Mariotti R
author_facet Busato A
Bonafede R
Bontempi P
Scambi I
Schiaffino L
Benati D
Malatesta M
Sbarbati A
Marzola P
Mariotti R
author_sort Busato A
title Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes
title_short Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes
title_full Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes
title_fullStr Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes
title_full_unstemmed Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes
title_sort magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes
publisher Dove Medical Press
publishDate 2016
url https://doaj.org/article/88895595986b40bbb3bb87ba84d3d04c
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