Cargando…

Culture Model for Non-human Primate Choroid Plexus

While there are murine and rat choroid plexus epithelial cell cultures, a translationally relevant model for choroid plexus activation and function is still lacking. The rhesus macaque is the gold standard for modeling viral infection and activation of CNS, including HIV-associated neurocognitive di...

Descripción completa

Detalles Bibliográficos
Autores principales: Delery, Elizabeth C., MacLean, Andrew G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6724611/
https://www.ncbi.nlm.nih.gov/pubmed/31555096
http://dx.doi.org/10.3389/fncel.2019.00396
_version_ 1783449027974529024
author Delery, Elizabeth C.
MacLean, Andrew G.
author_facet Delery, Elizabeth C.
MacLean, Andrew G.
author_sort Delery, Elizabeth C.
collection PubMed
description While there are murine and rat choroid plexus epithelial cell cultures, a translationally relevant model for choroid plexus activation and function is still lacking. The rhesus macaque is the gold standard for modeling viral infection and activation of CNS, including HIV-associated neurocognitive disorders. We have developed a rhesus macaque choroid plexus epithelial cell culture model which we believe to be suitable for studies of inflammation associated with viral infection of the CNS. Epithelial morphology and function were assessed using vimentin, phalloidin, the tight junction protein zonula-occludens-1 (ZO-1), and focal adhesion kinase (FAK). Choroid plexus epithelial cell type was confirmed using immunofluorescence with two proteins highly expressed in the choroid plexus: transthyretin and α-klotho. Finally, barrier properties of the model were monitored using pro- and anti-inflammatory mediators (TNF-α, the TLR2 agonist PamCys3K, and dexamethasone). When pro-inflammatory TNF-α was added to the xCelligence wells, there was a decrease in barrier function, which decreased in a step-wise fashion with each additional administration. This barrier function was repaired upon addition of the steroid dexamethasone. The TLR2 agonist PAM3CysK increased barrier functions in TNF-α treated wells. We have presented a model of the blood-CSF barrier that will allow study into pro- and anti-inflammatory conditions in the brain, while simultaneously measuring real time changes to epithelial cells.
format Online
Article
Text
id pubmed-6724611
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-67246112019-09-25 Culture Model for Non-human Primate Choroid Plexus Delery, Elizabeth C. MacLean, Andrew G. Front Cell Neurosci Neuroscience While there are murine and rat choroid plexus epithelial cell cultures, a translationally relevant model for choroid plexus activation and function is still lacking. The rhesus macaque is the gold standard for modeling viral infection and activation of CNS, including HIV-associated neurocognitive disorders. We have developed a rhesus macaque choroid plexus epithelial cell culture model which we believe to be suitable for studies of inflammation associated with viral infection of the CNS. Epithelial morphology and function were assessed using vimentin, phalloidin, the tight junction protein zonula-occludens-1 (ZO-1), and focal adhesion kinase (FAK). Choroid plexus epithelial cell type was confirmed using immunofluorescence with two proteins highly expressed in the choroid plexus: transthyretin and α-klotho. Finally, barrier properties of the model were monitored using pro- and anti-inflammatory mediators (TNF-α, the TLR2 agonist PamCys3K, and dexamethasone). When pro-inflammatory TNF-α was added to the xCelligence wells, there was a decrease in barrier function, which decreased in a step-wise fashion with each additional administration. This barrier function was repaired upon addition of the steroid dexamethasone. The TLR2 agonist PAM3CysK increased barrier functions in TNF-α treated wells. We have presented a model of the blood-CSF barrier that will allow study into pro- and anti-inflammatory conditions in the brain, while simultaneously measuring real time changes to epithelial cells. Frontiers Media S.A. 2019-08-28 /pmc/articles/PMC6724611/ /pubmed/31555096 http://dx.doi.org/10.3389/fncel.2019.00396 Text en Copyright © 2019 Delery and MacLean. 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 Neuroscience
Delery, Elizabeth C.
MacLean, Andrew G.
Culture Model for Non-human Primate Choroid Plexus
title Culture Model for Non-human Primate Choroid Plexus
title_full Culture Model for Non-human Primate Choroid Plexus
title_fullStr Culture Model for Non-human Primate Choroid Plexus
title_full_unstemmed Culture Model for Non-human Primate Choroid Plexus
title_short Culture Model for Non-human Primate Choroid Plexus
title_sort culture model for non-human primate choroid plexus
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6724611/
https://www.ncbi.nlm.nih.gov/pubmed/31555096
http://dx.doi.org/10.3389/fncel.2019.00396
work_keys_str_mv AT deleryelizabethc culturemodelfornonhumanprimatechoroidplexus
AT macleanandrewg culturemodelfornonhumanprimatechoroidplexus