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 & TRADE; (TC), (2) TC coated with the solubilized basement membrane matrix Matrigel(& REG;) (MM), (3) TC with the rainbow trout fibroblast cell line RTskin01 embedded within the Matrigel(& REG;) matrix (MMfb), or (4) the highly porous polystyrene scaffold Alvetex(& REG;) 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.

Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro / N. Verdile, F. Camin, R. Pavlovic, R. Pasquariello, M. Stuknytė, I. DE NONI, T.A.L. Brevini, F. Gandolfi. - In: CELLS. - ISSN 2073-4409. - 12:14(2023 Jul 13), pp. 1843.1-1843.26. [10.3390/cells12141843]

Distinct Organotypic Platforms Modulate Rainbow Trout (Oncorhynchus mykiss) Intestinal Cell Differentiation In Vitro

N. Verdile
Primo
Investigation
;
F. Camin
Secondo
Investigation
;
R. Pavlovic
Investigation
;
R. Pasquariello
Investigation
;
I. DE NONI
Conceptualization
;
T.A.L. Brevini
Penultimo
Conceptualization
;
F. Gandolfi
Ultimo
Writing – Review & Editing
2023

Abstract

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 & TRADE; (TC), (2) TC coated with the solubilized basement membrane matrix Matrigel(& REG;) (MM), (3) TC with the rainbow trout fibroblast cell line RTskin01 embedded within the Matrigel(& REG;) matrix (MMfb), or (4) the highly porous polystyrene scaffold Alvetex(& REG;) 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.
aquaculture; cell differentiation; enterocytes; in vitro intestine; intestinal cells; organoids; organotypic culture; rainbow trout; sustainable feed
Settore VET/01 - Anatomia degli Animali Domestici
Settore AGR/20 - Zoocolture
   Developing an Artificial Intestine for the sustainable farming of healthy fish (Fish-AI)
   Fish-AI
   EUROPEAN COMMISSION
   H2020
   828835
13-lug-2023
https://www.mdpi.com/2073-4409/12/14/1843
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/996297
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