Please use this identifier to cite or link to this item: http://hdl.handle.net/10316/95895
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dc.contributor.authorPires, Cristiana L.-
dc.contributor.authorPraça, Catarina-
dc.contributor.authorMartins, Patrícia A. T.-
dc.contributor.authorBatista de Carvalho, Ana L. M.-
dc.contributor.authorFerreira, Lino-
dc.contributor.authorCatarro, Maria Paula M. Marques-
dc.contributor.authorMoreno, Maria João-
dc.date.accessioned2021-10-12T10:50:40Z-
dc.date.available2021-10-12T10:50:40Z-
dc.date.issued2021-
dc.identifier.issn1999-4923pt
dc.identifier.urihttp://hdl.handle.net/10316/95895-
dc.description.abstractCaco-2 monolayers are a common in vitro model used to evaluate human intestinal absorption. The reference protocol requires 21 days post-seeding to establish a stable and confluent cell monolayer, which is used in a single permeability assay during the period of monolayer stability (up to day 30). In this work, we characterize variations in the tightness of the cell monolayer over the stable time interval and evaluate the conditions required for their re-use in permeability assays. The monolayer integrity was assessed through TEER measurements and permeability of the paracellular marker Lucifer Yellow (LY), complemented with nuclei and ZO-1 staining for morphological studies and the presence of tight junctions. Over 150 permeability assays were performed, which showed that manipulation of the cell monolayer in the permeability assay may contribute significantly to the flux of LY, leading to Papp values that are dependent on the sampling duration. The assay also leads to a small decrease in the cell monolayer TEER, which is fully recovered when cell monolayers are incubated with culture media for two full days. When this procedure is followed, the cell monolayers may be used for permeability assays on days 22, 25, and 28, triplicating the throughput of this important assay. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.pt
dc.language.isoengpt
dc.publisherMDPIpt
dc.relationUIDB/00070/2020pt
dc.relationNEUROATLANTIC/EAPA_791/2018pt
dc.relation2IQBIONEURO/0624_2IQBIONEURO_6_Ept
dc.relationSFRH/BD/138873/2018pt
dc.relationUIDB/00313/2020pt
dc.relationUIDP/00313/2020pt
dc.relationPT2020-PTDC/QUI-OUT/29373/2017pt
dc.relationPT2020-PTDC/DTP-FTO/2784/2014pt
dc.rightsopenAccesspt
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt
dc.subjectCaco-2 cellspt
dc.subjectHigh-throughputpt
dc.subjectIntestinal permeabilitypt
dc.subjectLucifer yellowpt
dc.subjectTight junctionspt
dc.titleRe-Use of Caco-2 Monolayers in Permeability Assays—Validation Regarding Cell Monolayer Integritypt
dc.typearticle-
degois.publication.firstPage1563pt
degois.publication.issue10pt
degois.publication.titlePharmaceuticspt
dc.peerreviewedyespt
dc.identifier.doi10.3390/pharmaceutics13101563pt
degois.publication.volume13pt
dc.date.embargo2021-01-01*
uc.date.periodoEmbargo0pt
item.languageiso639-1en-
item.fulltextCom Texto completo-
item.grantfulltextopen-
crisitem.author.deptFaculty of Medicine-
crisitem.author.deptFaculty of Sciences and Technology-
crisitem.author.parentdeptUniversity of Coimbra-
crisitem.author.parentdeptUniversity of Coimbra-
crisitem.author.researchunitCNC - Center for Neuroscience and Cell Biology-
crisitem.author.researchunitQFM-UC – Molecular Physical-Chemistry R&D Unit-
crisitem.author.orcid0000-0001-8985-9302-
crisitem.author.orcid0000-0002-8391-0055-
Appears in Collections:FCTUC Ciências da Vida - Artigos em Revistas Internacionais
FMUC Medicina - Artigos em Revistas Internacionais
I&D QFM-UC - Artigos em Revistas Internacionais
I&D CNC - Artigos em Revistas Internacionais
I&D CQC - Artigos em Revistas Internacionais
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