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A Preclinical Model to Assess Intestinal Barrier Integrity Using Canine Enteroids and Colonoids

MP. Corbett, V. Gabriel, V. Livania, D. Díaz-Regañón, A. Ralston, C. Zdyrski, D. Liu, S. Minkler, H. Wickham, A. Lincoln, K. Paukner, T. Atherly, MM. Merodio, DK. Sahoo, DK. Meyerholz, K. Allenspach, JP. Mochel

. 2025 ; 14 (3) : . [pub] 20250306

Status not-indexed Language English Country Switzerland

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc25008263

Grant support
1912948 U.S. National Science Foundation
FSU_0000022 ISU Startup

While two-dimensional (2D) cell cultures, such as Caco-2 and Madin-Darby canine kidney (MDCK) cells are widely used in a variety of biological models, these two-dimensional in vitro systems present inherent limitations in replicating the complexities of in vivo biology. Recent progress in three-dimensional organoid technology has the potential to address these limitations. In this study, the characteristics of conventional 2D cell culture systems were compared to those of canine intestinal organoids (enteroids, ENT, and colonoids, COL). Light microscopy and transmission electron microscopy were employed to evaluate the microanatomy of ENT, COL, Caco-2, and MDCK cell monolayers, while transepithelial electrical resistance (TEER) values were measured to assess monolayer integrity. The TEER values of canine ENT monolayers more closely approximated reported TEER values for human small intestines compared to Caco-2 and MDCK monolayers. Additionally, canine ENT demonstrated greater monolayer stability than Caco-2 and MDCK cells. Notably, while all systems displayed desmosomes, canine ENT and COL exclusively produced mucus. These findings highlight the potential of the canine organoid system as a more biologically relevant model for in vitro studies, addressing the limitations of conventional 2D cell culture systems.

References provided by Crossref.org

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$a While two-dimensional (2D) cell cultures, such as Caco-2 and Madin-Darby canine kidney (MDCK) cells are widely used in a variety of biological models, these two-dimensional in vitro systems present inherent limitations in replicating the complexities of in vivo biology. Recent progress in three-dimensional organoid technology has the potential to address these limitations. In this study, the characteristics of conventional 2D cell culture systems were compared to those of canine intestinal organoids (enteroids, ENT, and colonoids, COL). Light microscopy and transmission electron microscopy were employed to evaluate the microanatomy of ENT, COL, Caco-2, and MDCK cell monolayers, while transepithelial electrical resistance (TEER) values were measured to assess monolayer integrity. The TEER values of canine ENT monolayers more closely approximated reported TEER values for human small intestines compared to Caco-2 and MDCK monolayers. Additionally, canine ENT demonstrated greater monolayer stability than Caco-2 and MDCK cells. Notably, while all systems displayed desmosomes, canine ENT and COL exclusively produced mucus. These findings highlight the potential of the canine organoid system as a more biologically relevant model for in vitro studies, addressing the limitations of conventional 2D cell culture systems.
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$a Gabriel, Vojtech $u Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA $1 https://orcid.org/0000000327042297
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$a Livania, Vanessa $u Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA
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$a Ralston, Abigail $u 3D Health Solutions Inc., Athens, GA 30602, USA $1 https://orcid.org/0000000220718349
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$a Liu, Dongjie $u Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA
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$a Atherly, Todd $u Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA
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