Cargando…

Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability

The intestinal epithelial Caco-2 cell monolayer is a well-established in vitro model useful for predicting intestinal drug absorption in humans. Coculture models of Caco-2 and goblet-cell-like HT29-MTX cells have been developed to overcome the lack of a mucus layer; however, those models are much le...

Descripción completa

Detalles Bibliográficos
Autores principales: Cheng, Yi, Watanabe, Chie, Ando, Yusuke, Kitaoka, Satoshi, Egawa, Yuya, Takashima, Tomoya, Matsumoto, Akihiro, Murakami, Masahiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10535880/
https://www.ncbi.nlm.nih.gov/pubmed/37765306
http://dx.doi.org/10.3390/pharmaceutics15092338
_version_ 1785112734735532032
author Cheng, Yi
Watanabe, Chie
Ando, Yusuke
Kitaoka, Satoshi
Egawa, Yuya
Takashima, Tomoya
Matsumoto, Akihiro
Murakami, Masahiro
author_facet Cheng, Yi
Watanabe, Chie
Ando, Yusuke
Kitaoka, Satoshi
Egawa, Yuya
Takashima, Tomoya
Matsumoto, Akihiro
Murakami, Masahiro
author_sort Cheng, Yi
collection PubMed
description The intestinal epithelial Caco-2 cell monolayer is a well-established in vitro model useful for predicting intestinal drug absorption in humans. Coculture models of Caco-2 and goblet-cell-like HT29-MTX cells have been developed to overcome the lack of a mucus layer; however, those models are much leakier compared to the intestinal epithelium. Here, we developed a partially laminated culture model where HT29-MTX cells were superimposed onto a Caco-2 monolayer to overcome this issue. A morphological study showed that the piled HT29-MTX cells were voluntarily incorporated into the Caco-2 monolayer, and mucus production was confirmed via periodic acid-Schiff and mucin protein 2 staining. Permeability was evaluated in terms of transepithelial electrical resistance (TEER) and the apparent permeability of paracellular markers with different molecular sizes. The partially laminated model maintained the high barrier function of the Caco-2 monolayer, whose permeability appeared adjustable according to the HT29-MTX/Caco-2 cell ratio. In contrast, the coculture models showed abnormally high permeability of those markers, correlated with low TEER. Thus, the partially laminated model enabled in vitro recapitulation of effective mucosal barrier function. Consequently, this novel model may be useful as an in vitro high-throughput evaluation system for enteral mucosal permeability and mucus-penetrating efficiency of drugs and nanocarriers.
format Online
Article
Text
id pubmed-10535880
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-105358802023-09-29 Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability Cheng, Yi Watanabe, Chie Ando, Yusuke Kitaoka, Satoshi Egawa, Yuya Takashima, Tomoya Matsumoto, Akihiro Murakami, Masahiro Pharmaceutics Article The intestinal epithelial Caco-2 cell monolayer is a well-established in vitro model useful for predicting intestinal drug absorption in humans. Coculture models of Caco-2 and goblet-cell-like HT29-MTX cells have been developed to overcome the lack of a mucus layer; however, those models are much leakier compared to the intestinal epithelium. Here, we developed a partially laminated culture model where HT29-MTX cells were superimposed onto a Caco-2 monolayer to overcome this issue. A morphological study showed that the piled HT29-MTX cells were voluntarily incorporated into the Caco-2 monolayer, and mucus production was confirmed via periodic acid-Schiff and mucin protein 2 staining. Permeability was evaluated in terms of transepithelial electrical resistance (TEER) and the apparent permeability of paracellular markers with different molecular sizes. The partially laminated model maintained the high barrier function of the Caco-2 monolayer, whose permeability appeared adjustable according to the HT29-MTX/Caco-2 cell ratio. In contrast, the coculture models showed abnormally high permeability of those markers, correlated with low TEER. Thus, the partially laminated model enabled in vitro recapitulation of effective mucosal barrier function. Consequently, this novel model may be useful as an in vitro high-throughput evaluation system for enteral mucosal permeability and mucus-penetrating efficiency of drugs and nanocarriers. MDPI 2023-09-18 /pmc/articles/PMC10535880/ /pubmed/37765306 http://dx.doi.org/10.3390/pharmaceutics15092338 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cheng, Yi
Watanabe, Chie
Ando, Yusuke
Kitaoka, Satoshi
Egawa, Yuya
Takashima, Tomoya
Matsumoto, Akihiro
Murakami, Masahiro
Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability
title Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability
title_full Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability
title_fullStr Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability
title_full_unstemmed Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability
title_short Caco-2 Cell Sheet Partially Laminated with HT29-MTX Cells as a Novel In Vitro Model of Gut Epithelium Drug Permeability
title_sort caco-2 cell sheet partially laminated with ht29-mtx cells as a novel in vitro model of gut epithelium drug permeability
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10535880/
https://www.ncbi.nlm.nih.gov/pubmed/37765306
http://dx.doi.org/10.3390/pharmaceutics15092338
work_keys_str_mv AT chengyi caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability
AT watanabechie caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability
AT andoyusuke caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability
AT kitaokasatoshi caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability
AT egawayuya caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability
AT takashimatomoya caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability
AT matsumotoakihiro caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability
AT murakamimasahiro caco2cellsheetpartiallylaminatedwithht29mtxcellsasanovelinvitromodelofgutepitheliumdrugpermeability