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A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease

Glial cells are essential to understand Alzheimer’s disease (AD) progression, given their role in neuroinflammation and neurodegeneration. There is a need for reliable and easy to manipulate models that allow studying the mechanisms behind neuron and glia communication. Currently available models su...

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Autores principales: Luchena, Celia, Zuazo-Ibarra, Jone, Valero, Jorge, Matute, Carlos, Alberdi, Elena, Capetillo-Zarate, Estibaliz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9048896/
https://www.ncbi.nlm.nih.gov/pubmed/35493929
http://dx.doi.org/10.3389/fnagi.2022.844534
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author Luchena, Celia
Zuazo-Ibarra, Jone
Valero, Jorge
Matute, Carlos
Alberdi, Elena
Capetillo-Zarate, Estibaliz
author_facet Luchena, Celia
Zuazo-Ibarra, Jone
Valero, Jorge
Matute, Carlos
Alberdi, Elena
Capetillo-Zarate, Estibaliz
author_sort Luchena, Celia
collection PubMed
description Glial cells are essential to understand Alzheimer’s disease (AD) progression, given their role in neuroinflammation and neurodegeneration. There is a need for reliable and easy to manipulate models that allow studying the mechanisms behind neuron and glia communication. Currently available models such as co-cultures require complex methodologies and/or might not be affordable for all laboratories. With this in mind, we aimed to establish a straightforward in vitro setting with neurons and glial cells to study AD. We generated and optimized a 2D triple co-culture model with murine astrocytes, neurons and microglia, based on sequential seeding of each cell type. Immunofluorescence, western blot and ELISA techniques were used to characterize the effects of oligomeric Aβ (oAβ) in this model. We found that, in the triple co-culture, microglia increased the expression of anti-inflammatory marker Arginase I, and reduced pro-inflammatory iNOS and IL-1β, compared with microglia alone. Astrocytes reduced expression of pro-inflammatory A1 markers AMIGO2 and C3, and displayed a ramified morphology resembling physiological conditions. Anti-inflammatory marker TGF-β1 was also increased in the triple co-culture. Lastly, neurons increased post-synaptic markers, and developed more and longer branches than in individual primary cultures. Addition of oAβ in the triple co-culture reduced synaptic markers and increased CD11b in microglia, which are hallmarks of AD. Consequently, we developed a straightforward and reproducible triple co-cultured model, where cells resemble physiological conditions better than in individual primary cultures: microglia are less inflammatory, astrocytes are less reactive and neurons display a more mature morphology. Moreover, we are able to recapitulate Aβ-induced synaptic loss and CD11b increase. This model emerges as a powerful tool to study neurodegeneration and neuroinflammation in the context of AD and other neurodegenerative diseases.
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spelling pubmed-90488962022-04-29 A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease Luchena, Celia Zuazo-Ibarra, Jone Valero, Jorge Matute, Carlos Alberdi, Elena Capetillo-Zarate, Estibaliz Front Aging Neurosci Neuroscience Glial cells are essential to understand Alzheimer’s disease (AD) progression, given their role in neuroinflammation and neurodegeneration. There is a need for reliable and easy to manipulate models that allow studying the mechanisms behind neuron and glia communication. Currently available models such as co-cultures require complex methodologies and/or might not be affordable for all laboratories. With this in mind, we aimed to establish a straightforward in vitro setting with neurons and glial cells to study AD. We generated and optimized a 2D triple co-culture model with murine astrocytes, neurons and microglia, based on sequential seeding of each cell type. Immunofluorescence, western blot and ELISA techniques were used to characterize the effects of oligomeric Aβ (oAβ) in this model. We found that, in the triple co-culture, microglia increased the expression of anti-inflammatory marker Arginase I, and reduced pro-inflammatory iNOS and IL-1β, compared with microglia alone. Astrocytes reduced expression of pro-inflammatory A1 markers AMIGO2 and C3, and displayed a ramified morphology resembling physiological conditions. Anti-inflammatory marker TGF-β1 was also increased in the triple co-culture. Lastly, neurons increased post-synaptic markers, and developed more and longer branches than in individual primary cultures. Addition of oAβ in the triple co-culture reduced synaptic markers and increased CD11b in microglia, which are hallmarks of AD. Consequently, we developed a straightforward and reproducible triple co-cultured model, where cells resemble physiological conditions better than in individual primary cultures: microglia are less inflammatory, astrocytes are less reactive and neurons display a more mature morphology. Moreover, we are able to recapitulate Aβ-induced synaptic loss and CD11b increase. This model emerges as a powerful tool to study neurodegeneration and neuroinflammation in the context of AD and other neurodegenerative diseases. Frontiers Media S.A. 2022-04-14 /pmc/articles/PMC9048896/ /pubmed/35493929 http://dx.doi.org/10.3389/fnagi.2022.844534 Text en Copyright © 2022 Luchena, Zuazo-Ibarra, Valero, Matute, Alberdi and Capetillo-Zarate. https://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 Neuroscience
Luchena, Celia
Zuazo-Ibarra, Jone
Valero, Jorge
Matute, Carlos
Alberdi, Elena
Capetillo-Zarate, Estibaliz
A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease
title A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease
title_full A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease
title_fullStr A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease
title_full_unstemmed A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease
title_short A Neuron, Microglia, and Astrocyte Triple Co-culture Model to Study Alzheimer’s Disease
title_sort neuron, microglia, and astrocyte triple co-culture model to study alzheimer’s disease
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9048896/
https://www.ncbi.nlm.nih.gov/pubmed/35493929
http://dx.doi.org/10.3389/fnagi.2022.844534
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