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Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro
In vitro organotypic cell-based intestinal platforms, able to faithfully recapitulate the complex functions of the organ in vivo, would be a great support to search for more sustainable feed ingredients in aquaculture. We previously demonstrated that proliferation or differentiation of rainbow trout...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10377977/ https://www.ncbi.nlm.nih.gov/pubmed/37508507 http://dx.doi.org/10.3390/cells12141843 |
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author | Verdile, Nicole Camin, Federica Pavlovic, Radmila Pasquariello, Rolando Stuknytė, Milda De Noni, Ivano Brevini, Tiziana A. L. Gandolfi, Fulvio |
author_facet | Verdile, Nicole Camin, Federica Pavlovic, Radmila Pasquariello, Rolando Stuknytė, Milda De Noni, Ivano Brevini, Tiziana A. L. Gandolfi, Fulvio |
author_sort | Verdile, Nicole |
collection | PubMed |
description | In vitro organotypic cell-based intestinal platforms, able to faithfully recapitulate the complex functions of the organ in vivo, would be a great support to search for more sustainable feed ingredients in aquaculture. We previously demonstrated that proliferation or differentiation of rainbow trout intestinal cell lines is dictated by the culture environment. The aim of the present work was to develop a culture platform that can efficiently promote cell differentiation into mature enterocytes. We compared four options, seeding the RTpiMI cell line derived from the proximal intestine on (1) polyethylene terephthalate (PET) culture inserts ThinCert™ (TC), (2) TC coated with the solubilized basement membrane matrix Matrigel(®) (MM), (3) TC with the rainbow trout fibroblast cell line RTskin01 embedded within the Matrigel(®) matrix (MMfb), or (4) the highly porous polystyrene scaffold Alvetex(®) populated with the abovementioned fibroblast cell line (AV). We evaluated the presence of columnar cells with a clear polarization of brush border enzymes, the formation of an efficient barrier with a significant increase in transepithelial electrical resistance (TEER), and its ability to prevent the paracellular flux of large molecules but allow the transit of small compounds (proline and glucose) from the apical to the basolateral compartment. All parameters improved moving from the simplest (TC) through the more complex platforms. The presence of fibroblasts was particularly effective in enhancing epithelial cell differentiation within the AV platform recreating more closely the complexity of the intestinal mucosa, including the presence of extracellular vesicles between fibroblasts and epithelial cells. |
format | Online Article Text |
id | pubmed-10377977 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-103779772023-07-29 Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro Verdile, Nicole Camin, Federica Pavlovic, Radmila Pasquariello, Rolando Stuknytė, Milda De Noni, Ivano Brevini, Tiziana A. L. Gandolfi, Fulvio Cells Article In vitro organotypic cell-based intestinal platforms, able to faithfully recapitulate the complex functions of the organ in vivo, would be a great support to search for more sustainable feed ingredients in aquaculture. We previously demonstrated that proliferation or differentiation of rainbow trout intestinal cell lines is dictated by the culture environment. The aim of the present work was to develop a culture platform that can efficiently promote cell differentiation into mature enterocytes. We compared four options, seeding the RTpiMI cell line derived from the proximal intestine on (1) polyethylene terephthalate (PET) culture inserts ThinCert™ (TC), (2) TC coated with the solubilized basement membrane matrix Matrigel(®) (MM), (3) TC with the rainbow trout fibroblast cell line RTskin01 embedded within the Matrigel(®) matrix (MMfb), or (4) the highly porous polystyrene scaffold Alvetex(®) populated with the abovementioned fibroblast cell line (AV). We evaluated the presence of columnar cells with a clear polarization of brush border enzymes, the formation of an efficient barrier with a significant increase in transepithelial electrical resistance (TEER), and its ability to prevent the paracellular flux of large molecules but allow the transit of small compounds (proline and glucose) from the apical to the basolateral compartment. All parameters improved moving from the simplest (TC) through the more complex platforms. The presence of fibroblasts was particularly effective in enhancing epithelial cell differentiation within the AV platform recreating more closely the complexity of the intestinal mucosa, including the presence of extracellular vesicles between fibroblasts and epithelial cells. MDPI 2023-07-13 /pmc/articles/PMC10377977/ /pubmed/37508507 http://dx.doi.org/10.3390/cells12141843 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 Verdile, Nicole Camin, Federica Pavlovic, Radmila Pasquariello, Rolando Stuknytė, Milda De Noni, Ivano Brevini, Tiziana A. L. Gandolfi, Fulvio Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro |
title | Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro |
title_full | Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro |
title_fullStr | Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro |
title_full_unstemmed | Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro |
title_short | Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro |
title_sort | distinct organotypic platforms modulate rainbow trout (oncorhynchus mykiss) intestinal cell differentiation in vitro |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10377977/ https://www.ncbi.nlm.nih.gov/pubmed/37508507 http://dx.doi.org/10.3390/cells12141843 |
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