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...

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Main Authors: Elizabeth C. Delery, Andrew G. MacLean
Format: Article
Language:English
Published: Frontiers Media S.A. 2019-08-01
Series:Frontiers in Cellular Neuroscience
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fncel.2019.00396/full
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spelling doaj-9892cea6324f4b25b8da6d82e87b3e0f2020-11-24T21:21:38ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022019-08-011310.3389/fncel.2019.00396479548Culture Model for Non-human Primate Choroid PlexusElizabeth C. Delery0Elizabeth C. Delery1Elizabeth C. Delery2Andrew G. MacLean3Andrew G. MacLean4Andrew G. MacLean5Andrew G. MacLean6Andrew G. MacLean7Division of Comparative Pathology, Tulane National Primate Research Center, Covington, LA, United StatesTulane Program in Biomedical Sciences, New Orleans, LA, United StatesDepartment of Microbiology and Immunology, Tulane Medical School, New Orleans, LA, United StatesDivision of Comparative Pathology, Tulane National Primate Research Center, Covington, LA, United StatesTulane Program in Biomedical Sciences, New Orleans, LA, United StatesDepartment of Microbiology and Immunology, Tulane Medical School, New Orleans, LA, United StatesTulane Brain Institute, New Orleans, LA, United StatesTulane Center for Aging, New Orleans, LA, United StatesWhile 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.https://www.frontiersin.org/article/10.3389/fncel.2019.00396/fullchoroid plexusepithelial cellcell culturerhesus macaqueaginginfectious disease
collection DOAJ
language English
format Article
sources DOAJ
author Elizabeth C. Delery
Elizabeth C. Delery
Elizabeth C. Delery
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
spellingShingle Elizabeth C. Delery
Elizabeth C. Delery
Elizabeth C. Delery
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
Culture Model for Non-human Primate Choroid Plexus
Frontiers in Cellular Neuroscience
choroid plexus
epithelial cell
cell culture
rhesus macaque
aging
infectious disease
author_facet Elizabeth C. Delery
Elizabeth C. Delery
Elizabeth C. Delery
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
Andrew G. MacLean
author_sort Elizabeth C. Delery
title Culture Model for Non-human Primate Choroid Plexus
title_short 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_sort culture model for non-human primate choroid plexus
publisher Frontiers Media S.A.
series Frontiers in Cellular Neuroscience
issn 1662-5102
publishDate 2019-08-01
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.
topic choroid plexus
epithelial cell
cell culture
rhesus macaque
aging
infectious disease
url https://www.frontiersin.org/article/10.3389/fncel.2019.00396/full
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