A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium
Abstract Epithelial damage due to gastrointestinal disorders frequently causes severe disease in horses. To study the underlying pathophysiological processes, we aimed to establish equine jejunum and colon enteroids (eqJE, eqCE) mimicking the in vivo epithelium. Therefore, enteroids were cultivated...
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s13567-024-01283-0 |
| _version_ | 1827315870862409728 |
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| author | Christina Windhaber Anna Heckl Georg Csukovich Barbara Pratscher Iwan Anton Burgener Nora Biermann Franziska Dengler |
| author_facet | Christina Windhaber Anna Heckl Georg Csukovich Barbara Pratscher Iwan Anton Burgener Nora Biermann Franziska Dengler |
| author_sort | Christina Windhaber |
| collection | DOAJ |
| description | Abstract Epithelial damage due to gastrointestinal disorders frequently causes severe disease in horses. To study the underlying pathophysiological processes, we aimed to establish equine jejunum and colon enteroids (eqJE, eqCE) mimicking the in vivo epithelium. Therefore, enteroids were cultivated in four different media for differentiation and subsequently characterized histomorphologically, on mRNA and on protein level in comparison to the native epithelium of the same donor horses to identify ideal culture conditions for an in vitro model system. With increasing enterocyte differentiation, the enteroids showed a reduced growth rate as well as a predominantly spherical morphology and less budding compared to enteroids in proliferation medium. Combined or individual withdrawal of stem cell niche pathway components resulted in lower mRNA expression levels of stem cell markers and concomitant differentiation of enterocytes, goblet cells and enteroendocrine cells. For eqCE, withdrawal of Wnt alone was sufficient for the generation of differentiated enterocytes with a close resemblance to the in vivo epithelium. Combined removal of Wnt, R-spondin and Noggin and the addition of DAPT stimulated differentiation of eqJE at a similar level as the in vivo epithelium, particularly with regard to enterocytes. In summary, we successfully defined a medium composition that promotes the formation of eqJE and eqCE consisting of multiple cell types and resembling the in vivo epithelium. Our findings emphasize the importance of adapting culture conditions to the respective species and the intestinal segment. This in vitro model will be used to investigate the pathological mechanisms underlying equine gastrointestinal disorders in future studies. |
| first_indexed | 2024-04-24T23:04:09Z |
| format | Article |
| id | doaj.art-cbc7698439954805aa906a3c50642792 |
| institution | Directory Open Access Journal |
| issn | 1297-9716 |
| language | English |
| last_indexed | 2024-04-24T23:04:09Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj.art-cbc7698439954805aa906a3c506427922024-03-17T12:35:08ZengBMCVeterinary Research1297-97162024-03-0155112110.1186/s13567-024-01283-0A matter of differentiation: equine enteroids as a model for the in vivo intestinal epitheliumChristina Windhaber0Anna Heckl1Georg Csukovich2Barbara Pratscher3Iwan Anton Burgener4Nora Biermann5Franziska Dengler6Institute of Physiology, Pathophysiology and Biophysics, University of Veterinary MedicineInstitute of Physiology, Pathophysiology and Biophysics, University of Veterinary MedicineDivision of Small Animal Internal Medicine, University of Veterinary MedicineDivision of Small Animal Internal Medicine, University of Veterinary MedicineDivision of Small Animal Internal Medicine, University of Veterinary MedicineClinical Unit of Equine Surgery, University of Veterinary MedicineInstitute of Physiology, Pathophysiology and Biophysics, University of Veterinary MedicineAbstract Epithelial damage due to gastrointestinal disorders frequently causes severe disease in horses. To study the underlying pathophysiological processes, we aimed to establish equine jejunum and colon enteroids (eqJE, eqCE) mimicking the in vivo epithelium. Therefore, enteroids were cultivated in four different media for differentiation and subsequently characterized histomorphologically, on mRNA and on protein level in comparison to the native epithelium of the same donor horses to identify ideal culture conditions for an in vitro model system. With increasing enterocyte differentiation, the enteroids showed a reduced growth rate as well as a predominantly spherical morphology and less budding compared to enteroids in proliferation medium. Combined or individual withdrawal of stem cell niche pathway components resulted in lower mRNA expression levels of stem cell markers and concomitant differentiation of enterocytes, goblet cells and enteroendocrine cells. For eqCE, withdrawal of Wnt alone was sufficient for the generation of differentiated enterocytes with a close resemblance to the in vivo epithelium. Combined removal of Wnt, R-spondin and Noggin and the addition of DAPT stimulated differentiation of eqJE at a similar level as the in vivo epithelium, particularly with regard to enterocytes. In summary, we successfully defined a medium composition that promotes the formation of eqJE and eqCE consisting of multiple cell types and resembling the in vivo epithelium. Our findings emphasize the importance of adapting culture conditions to the respective species and the intestinal segment. This in vitro model will be used to investigate the pathological mechanisms underlying equine gastrointestinal disorders in future studies.https://doi.org/10.1186/s13567-024-01283-0Organoidjejunumcolonhorsethree-dimensional cell culturecolonoid |
| spellingShingle | Christina Windhaber Anna Heckl Georg Csukovich Barbara Pratscher Iwan Anton Burgener Nora Biermann Franziska Dengler A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium Veterinary Research Organoid jejunum colon horse three-dimensional cell culture colonoid |
| title | A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium |
| title_full | A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium |
| title_fullStr | A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium |
| title_full_unstemmed | A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium |
| title_short | A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium |
| title_sort | matter of differentiation equine enteroids as a model for the in vivo intestinal epithelium |
| topic | Organoid jejunum colon horse three-dimensional cell culture colonoid |
| url | https://doi.org/10.1186/s13567-024-01283-0 |
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