Cargando…

Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung

A successful clinical translation of novel nanoparticle-based cancer therapeutics requires a thorough preclinical investigation of their interaction with immune, tumor and endothelial cells as well as components of the tumor-microenvironment. Although high-resolution microscopy images of fixed tumor...

Descripción completa

Detalles Bibliográficos
Autores principales: Ramos-Gomes, Fernanda, Ferreira, Nathalia, Kraupner, Alexander, Alves, Frauke, Markus, M. Andrea
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7661932/
https://www.ncbi.nlm.nih.gov/pubmed/33195152
http://dx.doi.org/10.3389/fbioe.2020.588922
_version_ 1783609300850049024
author Ramos-Gomes, Fernanda
Ferreira, Nathalia
Kraupner, Alexander
Alves, Frauke
Markus, M. Andrea
author_facet Ramos-Gomes, Fernanda
Ferreira, Nathalia
Kraupner, Alexander
Alves, Frauke
Markus, M. Andrea
author_sort Ramos-Gomes, Fernanda
collection PubMed
description A successful clinical translation of novel nanoparticle-based cancer therapeutics requires a thorough preclinical investigation of their interaction with immune, tumor and endothelial cells as well as components of the tumor-microenvironment. Although high-resolution microscopy images of fixed tumor tissue specimens can provide valuable information in this regard, they are only static snapshots of a momentary event. Here we describe a superior alternative fluorescence microscopy approach to assess the feasibility of investigating nanoparticle-cell interactions in the mouse lung live and over time at nanometer resolution. We applied fluorescent lung tumor cells and Barium-based fluorescently labeled nanoparticles to nude mice or to CD68-EGFP transgenic mice for visualization of the monocyte-macrophage lineage. Shortly before imaging, fluorescently labeled lectin was intravenously injected for staining of the blood vessels. The lung was filled ex vivo with 1% agarose and individual lung lobes were imaged over time using a confocal microscope with Airyscan technology. Time series demonstrate that live cell imaging of lung lobes can be performed for at least 4 h post mortem. Time-lapse movies illustrate the dynamics of the nanoparticles within the pulmonary circulation and their uptake by immune cells. Moreover, the exchange of nanoparticle material between cancer cells was observed over time. Fluorescent monocytes in lungs of CD68-EGFP transgenic mice could be visualized within blood vessels in the process of interaction with tumor cells and nanoparticles. This high resolution ex vivo live cell imaging approach provides an excellent 4D tool to obtain valuable information on the behavior of tumor and immune cells at first encounter with nanoparticles and may contribute to the understanding of how nanoparticles interact with cells supporting the development of therapeutic strategies based on nanoparticulate drug delivery systems.
format Online
Article
Text
id pubmed-7661932
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-76619322020-11-13 Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung Ramos-Gomes, Fernanda Ferreira, Nathalia Kraupner, Alexander Alves, Frauke Markus, M. Andrea Front Bioeng Biotechnol Bioengineering and Biotechnology A successful clinical translation of novel nanoparticle-based cancer therapeutics requires a thorough preclinical investigation of their interaction with immune, tumor and endothelial cells as well as components of the tumor-microenvironment. Although high-resolution microscopy images of fixed tumor tissue specimens can provide valuable information in this regard, they are only static snapshots of a momentary event. Here we describe a superior alternative fluorescence microscopy approach to assess the feasibility of investigating nanoparticle-cell interactions in the mouse lung live and over time at nanometer resolution. We applied fluorescent lung tumor cells and Barium-based fluorescently labeled nanoparticles to nude mice or to CD68-EGFP transgenic mice for visualization of the monocyte-macrophage lineage. Shortly before imaging, fluorescently labeled lectin was intravenously injected for staining of the blood vessels. The lung was filled ex vivo with 1% agarose and individual lung lobes were imaged over time using a confocal microscope with Airyscan technology. Time series demonstrate that live cell imaging of lung lobes can be performed for at least 4 h post mortem. Time-lapse movies illustrate the dynamics of the nanoparticles within the pulmonary circulation and their uptake by immune cells. Moreover, the exchange of nanoparticle material between cancer cells was observed over time. Fluorescent monocytes in lungs of CD68-EGFP transgenic mice could be visualized within blood vessels in the process of interaction with tumor cells and nanoparticles. This high resolution ex vivo live cell imaging approach provides an excellent 4D tool to obtain valuable information on the behavior of tumor and immune cells at first encounter with nanoparticles and may contribute to the understanding of how nanoparticles interact with cells supporting the development of therapeutic strategies based on nanoparticulate drug delivery systems. Frontiers Media S.A. 2020-10-30 /pmc/articles/PMC7661932/ /pubmed/33195152 http://dx.doi.org/10.3389/fbioe.2020.588922 Text en Copyright © 2020 Ramos-Gomes, Ferreira, Kraupner, Alves and Markus. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Ramos-Gomes, Fernanda
Ferreira, Nathalia
Kraupner, Alexander
Alves, Frauke
Markus, M. Andrea
Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung
title Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung
title_full Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung
title_fullStr Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung
title_full_unstemmed Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung
title_short Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung
title_sort ex vivo live cell imaging of nanoparticle-cell interactions in the mouse lung
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7661932/
https://www.ncbi.nlm.nih.gov/pubmed/33195152
http://dx.doi.org/10.3389/fbioe.2020.588922
work_keys_str_mv AT ramosgomesfernanda exvivolivecellimagingofnanoparticlecellinteractionsinthemouselung
AT ferreiranathalia exvivolivecellimagingofnanoparticlecellinteractionsinthemouselung
AT kraupneralexander exvivolivecellimagingofnanoparticlecellinteractionsinthemouselung
AT alvesfrauke exvivolivecellimagingofnanoparticlecellinteractionsinthemouselung
AT markusmandrea exvivolivecellimagingofnanoparticlecellinteractionsinthemouselung