{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T05:18:57Z","timestamp":1788153537666,"version":"build-2803163510"},"reference-count":47,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:p>\n                    Choroid plexus epithelial cells (CPEC) are implicated in cerebrospinal fluid (CSF) production and the site of the inner blood-cerebrospinal fluid barrier. CPEC are specialized ependymal cells with characteristic cell junctions, intermediate filaments, and transport protein expression, including aquaporins. We have recently found that, in addition to aquaporin-1, a subset of CPEC in the human brain express aquaporin-4 (AQP4), the main water channel in the brain that has been implicated in water homeostasis and glymphatic system function. We therefore aimed to establish primary cultures from human CP to study AQP4 expression. We collected CP tissue from human body donors\n                    <jats:italic>post mortem<\/jats:italic>\n                    to grow CPEC\n                    <jats:italic>in vitro<\/jats:italic>\n                    . We successfully established two primary culture models for human CP tissue. In an organ culture model, small pieces of CP tissue were explanted on PTFE membranes. In a second model, cells from CP were dissociated and seeded on either laminin or collagen substrate. We monitored cell and tissue growth and investigated protein expression by immunofluorescence. Tissue integrity of explant cultures was maintained throughout the cultivation period of several weeks. Immunofluorescence signals for zonula occludens protein-1 (ZO1), AQP1, and Na\/K-ATPase were present although the expression patterns became increasingly irregular and non-polar over time. In dissociation cultures, cells attached and grew very slowly initially but reached confluency after several weeks. Cultures were passaged up to 6 times and could be cryopreserved. Application of cytosine arabinoside suppressed the growth of macrophages and most likely fibroblasts. These primary dissociation cultures were identified as epithelial by their morphology and positive immunofluorescence for transthyretin, vimentin. Na\/K-ATPase, ZO-1, cytokeratin, and AQP1. In both culture models, cells expressed AQP4. In dissociation cultures, AQP4 appeared to colocalize with ZO-1 at cell\u2013cell contact sites in many CPEC but showed separate localizations at high resolution. These studies show that primary organ and cell cultures can be generated from human\n                    <jats:italic>post mortem<\/jats:italic>\n                    CP tissue suitable to investigate regulation and localization of AQP4. Together, these culture models provide valuable tools to investigate the proteins involved in CSF production and glymphatic clearance, relevant for the pathogenesis of neurodegenerative diseases.\n                  <\/jats:p>","DOI":"10.3389\/fncel.2026.1827493","type":"journal-article","created":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T04:29:37Z","timestamp":1788150577000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Primary cell and tissue cultures of human choroid plexus epithelial cells"],"prefix":"10.3389","volume":"20","author":[{"given":"Ronja","family":"Bihlmaier","sequence":"first","affiliation":[{"name":"Institute of Clinical Anatomy and Cell Analysis, Eberhard Karls Universit\u00e4t T\u00fcbingen","place":["T\u00fcbingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melanie","family":"Scharr","sequence":"additional","affiliation":[{"name":"Institute of Clinical Anatomy and Cell Analysis, Eberhard Karls Universit\u00e4t T\u00fcbingen","place":["T\u00fcbingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter H.","family":"Neckel","sequence":"additional","affiliation":[{"name":"Institute of Clinical Anatomy and Cell Analysis, Eberhard Karls Universit\u00e4t T\u00fcbingen","place":["T\u00fcbingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernhard","family":"Hirt","sequence":"additional","affiliation":[{"name":"Institute of Clinical Anatomy and Cell Analysis, Eberhard Karls Universit\u00e4t T\u00fcbingen","place":["T\u00fcbingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas F.","family":"Mack","sequence":"additional","affiliation":[{"name":"Institute of Clinical Anatomy and Cell Analysis, Eberhard Karls Universit\u00e4t T\u00fcbingen","place":["T\u00fcbingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1038\/nrn1824","article-title":"Astrocyte-endothelial interactions at the blood-brain barrier","volume":"7","author":"Abbott","year":"2006","journal-title":"Nat. Rev. Neurosci."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"e0121738","DOI":"10.1371\/journal.pone.0121738","article-title":"Proliferation of cultured mouse choroid plexus epithelial cells","volume":"10","author":"Barkho","year":"2015","journal-title":"PLoS One"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"954","DOI":"10.1002\/glia.23941","article-title":"Dp71 contribution to the molecular scaffold anchoring aquaporine-4 channels in brain macroglial cells","volume":"69","author":"Belmaati Cherkaoui","year":"2021","journal-title":"Glia"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"2973","DOI":"10.1007\/s11095-015-1679-1","article-title":"Characterization of efflux transport proteins of the human choroid plexus papilloma cell line Hibcpp, a functional in vitro model of the blood-cerebrospinal fluid barrier","volume":"32","author":"Bernd","year":"2015","journal-title":"Pharm. Res."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"212","DOI":"10.3390\/biom13020212","article-title":"Aquaporin-1 and Aquaporin-4 expression in Ependyma, choroid plexus and surrounding transition zones in the human brain","volume":"13","author":"Bihlmaier","year":"2023","journal-title":"Biomolecules"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"2787","DOI":"10.1007\/s00018-013-1300-x","article-title":"Neurogenic effects of \u03b2-amyloid in the choroid plexus epithelial cells in Alzheimer\u2019s disease","volume":"70","author":"Bolos","year":"2013","journal-title":"Cell. Mol. Life Sci."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1111\/j.1471-4159.1981.tb04470.x","article-title":"Culture and characterization of epithelial cells from bovine choroid plexus","volume":"37","author":"Crook","year":"1981","journal-title":"J. Neurochem."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"3056","DOI":"10.1016\/j.cell.2021.04.003","article-title":"A cellular and spatial map of the choroid plexus across brain ventricles and ages","volume":"184","author":"Dani","year":"2021","journal-title":"Cell"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"90","DOI":"10.1007\/s00018-022-04136-1","article-title":"Aquaporin-4 expression in the human choroid plexus","volume":"79","author":"Deffner","year":"2022","journal-title":"Cell. Mol. Life Sci."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"396","DOI":"10.3389\/fncel.2019.00396","article-title":"Culture model for non-human primate choroid plexus","volume":"13","author":"Delery","year":"2019","journal-title":"Front. Cell. Neurosci."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"579","DOI":"10.1016\/j.it.2012.07.004","article-title":"Capture, crawl, cross: the T cell code to breach the blood-brain barriers","volume":"33","author":"Engelhardt","year":"2012","journal-title":"Trends Immunol."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"1141","DOI":"10.1046\/j.1471-4159.1998.71031141.x","article-title":"The polarity of choroid plexus epithelial cells in vitro is improved in serum free medium","volume":"71","author":"Hakvoort","year":"1998","journal-title":"J. Neurochem."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1002\/1097-0029(20010101)52:1<137::AID-JEMT15>3.0.CO;2-J","article-title":"Porcine choroid plexus epithelial cells in culture: regulation of barrier properties and transport processes","volume":"52","author":"Haselbach","year":"2001","journal-title":"Microsc. Res. Tech."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1111\/j.1749-0774.2005.tb00059.x","article-title":"Establishment and characterization of a human malignant choroids plexus papilloma cell line (Hibcpp)","volume":"18","author":"Ishiwata","year":"2005","journal-title":"Hum. Cell"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"547","DOI":"10.1038\/s41583-019-0201-x","article-title":"Macrophages at Cns interfaces: ontogeny and function in health and disease","volume":"20","author":"Kierdorf","year":"2019","journal-title":"Nat. Rev. Neurosci."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"364","DOI":"10.1002\/glia.1123","article-title":"Differentiation of choroid plexus ependymal cells into astrocytes after grafting into the pre-lesioned spinal cord in mice","volume":"36","author":"Kitada","year":"2001","journal-title":"Glia"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"784","DOI":"10.1016\/j.cell.2020.03.037","article-title":"Targeting Aquaporin-4 subcellular localization to treat central nervous system edema","volume":"181","author":"Kitchen","year":"2020","journal-title":"Cell"},{"key":"ref18","doi-asserted-by":"publisher","DOI":"10.21769\/BioProtoc.3532","article-title":"Method for primary epithelial cell culture from the rat choroid plexus","volume":"10","author":"Lallai","year":"2020","journal-title":"Bio-Protoc."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1186\/s12987-016-0027-0","article-title":"Modeling immune functions of the mouse blood-cerebrospinal fluid barrier in vitro: primary rather than immortalized mouse choroid plexus epithelial cells are suited to study immune cell migration across this brain barrier","volume":"13","author":"Lazarevic","year":"2016","journal-title":"Fluids Barriers CNS"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1016\/S0022-510X(01)00657-8","article-title":"Cell proliferation and differentiation from ependymal, subependymal and choroid plexus cells in response to stroke in rats","volume":"193","author":"Li","year":"2002","journal-title":"J. Neurol. Sci."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"32","DOI":"10.3389\/fnins.2015.00032","article-title":"Development of the choroid plexus and blood-Csf barrier","volume":"9","author":"Liddelow","year":"2015","journal-title":"Front. Neurosci."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1177\/1073858412447981","article-title":"A novel look at astrocytes: Aquaporins, ionic homeostasis, and the role of the microenvironment for regeneration in the Cns","volume":"19","author":"Mack","year":"2013","journal-title":"Neuroscientist"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"1743","DOI":"10.1007\/s10571-020-00944-w","article-title":"The laminin-induced phosphorylation of Pkc\u03b4 regulates Aqp4 distribution and water permeability in rat astrocytes","volume":"41","author":"No\u00ebl","year":"","journal-title":"Cell. Mol. Neurobiol."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1002\/cne.24763","article-title":"Agrin plays a major role in the coalescence of the aquaporin-4 clusters induced by gamma-1-containing laminin","volume":"528","author":"No\u00ebl","year":"","journal-title":"J. Comp. Neurol."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"2109","DOI":"10.1111\/j.1460-9568.2007.05850.x","article-title":"Effects of agrin on the expression and distribution of the water channel protein aquaporin-4 and volume regulation in cultured astrocytes","volume":"26","author":"Noell","year":"2007","journal-title":"Eur. J. Neurosci."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"eaaz5626","DOI":"10.1126\/science.aaz5626","article-title":"Human CNS barrier-forming organoids with cerebrospinal fluid production","volume":"369","author":"Pellegrini","year":"2020","journal-title":"Science"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1007\/BF01111935","article-title":"The surface fine structure of the choroid plexus and ependymal lining of the rat lateral ventricle","volume":"3","author":"Peters","year":"1974","journal-title":"J. Neurocytol."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"C673","DOI":"10.1152\/ajpcell.00041.2017","article-title":"Transport across the choroid plexus epithelium","volume":"312","author":"Praetorius","year":"2017","journal-title":"Am. J. Phys. Cell Phys."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"1025","DOI":"10.1152\/physrev.00031.2020","article-title":"Fluid transport in the brain","volume":"102","author":"Rasmussen","year":"2022","journal-title":"Physiol. Rev."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1186\/2045-8118-10-10","article-title":"Studies on the human choroid plexus in vitro","volume":"10","author":"Redzic","year":"2013","journal-title":"Fluids Barriers Cns"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"214","DOI":"10.1038\/nature09611","article-title":"Developmental genetics of vertebrate glial-cell specification","volume":"468","author":"Rowitch","year":"2010","journal-title":"Nature"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"e39835","DOI":"10.1371\/journal.pone.0039835","article-title":"A novel porcine in vitro model of the blood-cerebrospinal fluid barrier with strong barrier function","volume":"7","author":"Schroten","year":"2012","journal-title":"PLoS One"},{"key":"ref33","doi-asserted-by":"publisher","first-page":"S515","DOI":"10.2741\/1085","article-title":"Choroid plexus, ageing of the brain, and Alzheimer's disease","volume":"8","author":"Serot","year":"2003","journal-title":"Front. Biosci. (Landmark Ed)"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"794","DOI":"10.1046\/j.0953-816x.2001.01693.x","article-title":"Choroid plexus and ageing in rats: a morphometric and ultrastructural study","volume":"14","author":"Serot","year":"2001","journal-title":"Eur. J. Neurosci."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"2116","DOI":"10.1210\/endo.133.5.8404661","article-title":"Thyroxine transport to the brain: role of protein synthesis by the choroid plexus","volume":"133","author":"Southwell","year":"1993","journal-title":"Endocrinology"},{"key":"ref36","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1016\/j.expneurol.2015.02.032","article-title":"A balanced view of choroid plexus structure and function: focus on adult humans","volume":"267","author":"Spector","year":"2015","journal-title":"Exp. Neurol."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"104014","DOI":"10.1016\/j.isci.2022.104014","article-title":"Transmigration of Trypanosoma brucei across an in vitro blood-cerebrospinal fluid barrier","volume":"25","author":"Speidel","year":"2022","journal-title":"iScience"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"6275","DOI":"10.1523\/JNEUROSCI.19-15-06275.1999","article-title":"Demonstration of a coupled metabolism-efflux process at the choroid plexus as a mechanism of brain protection toward xenobiotics","volume":"19","author":"Strazielle","year":"1999","journal-title":"J. Neurosci."},{"key":"ref39","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1007\/978-1-61779-077-5_8","volume-title":"Cell Culture Techniques","author":"Strazielle","year":"2011"},{"key":"ref40","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1186\/1743-8454-4-11","article-title":"Expression of junctional proteins in choroid plexus epithelial cell lines: a comparative study","volume":"4","author":"Szmydynger-Chodobska","year":"2007","journal-title":"Cerebrospinal Fluid Res."},{"key":"ref41","doi-asserted-by":"publisher","first-page":"9626","DOI":"10.1016\/S0021-9258(18)60576-9","article-title":"Transferrin gene expression and synthesis by cultured choroid plexus epithelial cells: regulation by serotonin and cyclic adenosine 3\u2032,5\u2032-monophosphate","volume":"264","author":"Tsutsumi","year":"1989","journal-title":"J. Biol. Chem."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"1109","DOI":"10.1016\/S0022-3565(24)36887-9","article-title":"Functional characterization of choroid plexus epithelial cells in primary Culture1","volume":"282","author":"Villalobos","year":"1997","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref43","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1016\/S0006-8993(99)01466-3","article-title":"Biondi ring tangles in the choroid plexus of Alzheimer's disease and normal aging brains: a quantitative study","volume":"832","author":"Wen","year":"1999","journal-title":"Brain Res."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1007\/s00401-009-0627-8","article-title":"Choroid plexus: biology and pathology","volume":"119","author":"Wolburg","year":"2010","journal-title":"Acta Neuropathol."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1016\/S0304-3940(01)01927-9","article-title":"Claudin-1, claudin-2 and claudin-11 are present in tight junctions of choroid plexus epithelium of the mouse","volume":"307","author":"Wolburg","year":"2001","journal-title":"Neurosci. Lett."},{"key":"ref46","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1002\/jmv.1890080405","article-title":"Comparison of human cytomegalovirus growth in Mrc-5 human fibroblasts, brain, and choroid plexus cells in vitro","volume":"8","author":"Wroblewska","year":"1981","journal-title":"J. Med. Virol."},{"key":"ref47","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1007\/s11626-998-0051-8","article-title":"Primary culture of choroidal epithelial cells: characterization of an in vitro model of blood-Csf barrier","volume":"34","author":"Zheng","year":"1998","journal-title":"In Vitro Cell. Dev. Biol. - Anim."}],"container-title":["Frontiers in Cellular Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2026.1827493\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T04:29:39Z","timestamp":1788150579000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2026.1827493\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,31]]},"references-count":47,"alternative-id":["10.3389\/fncel.2026.1827493"],"URL":"https:\/\/doi.org\/10.3389\/fncel.2026.1827493","relation":{},"ISSN":["1662-5102"],"issn-type":[{"value":"1662-5102","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,31]]},"article-number":"1827493"}}