Initial pig developmental stage influences intestinal organoid growth but not cellular composition
Abstract Background Intestinal organoids are promising tools in the context of animal experiment reduction but a thorough characterization of the impact of the origin of intestinal stem cells (ISC) on organoid phenotype is needed to routinely use this cellular model. Our objective was to evaluate th...
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Wiley
2024-12-01
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| Series: | Animal Models and Experimental Medicine |
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| Online Access: | https://doi.org/10.1002/ame2.12512 |
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| author | Camille Duchesne Gwénaëlle Randuineau Laurence Le Normand Véronique Romé Samia Laraqui Alexis Pierre Arnaud Gaëlle Boudry |
| author_facet | Camille Duchesne Gwénaëlle Randuineau Laurence Le Normand Véronique Romé Samia Laraqui Alexis Pierre Arnaud Gaëlle Boudry |
| author_sort | Camille Duchesne |
| collection | DOAJ |
| description | Abstract Background Intestinal organoids are promising tools in the context of animal experiment reduction but a thorough characterization of the impact of the origin of intestinal stem cells (ISC) on organoid phenotype is needed to routinely use this cellular model. Our objective was to evaluate the effect of ISC donor age on the growth, morphology and cellular composition of intestinal organoids derived from pig. Methods Organoids were derived from jejunal and colonic ISC obtained from 1‐, 7‐, 28‐, 36‐ and 180‐day‐old pigs and passaged three times. Results We first confirmed by qPCR that the expression of 18% of the >80 studied genes related to various intestinal functions differed between jejunal and colonic organoids after two passages (p < 0.05). Growth and morphology of organoids depended on intestinal location (greater number and larger organoids derived from colonic than jejunal ISC, p < 0.05) but also pig age. Indeed, when ISC were derived from young piglets, the ratio of organoids to spheroids was greater (p < 0.05), spheroids were larger during the primary culture but smaller after two passages (p < 0.05) and organoids were smaller after one passage (p > 0.05) compared to ISC from older pigs. Finally, no difference in cellular composition, evaluated by immunostaining of markers of the major intestinal cell types (absorptive, enteroendocrine and goblet cells) was observed between organoids originating from 7‐ or 180‐day‐old pigs, but differences between intestinal site origins were noticed. Conclusion In conclusion, while the age of the tissue donor affected organoid growth and morphology, it did not influence the phenotype. |
| format | Article |
| id | doaj-art-40d37611aa6845deb5073c00bbd875ca |
| institution | OA Journals |
| issn | 2576-2095 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Animal Models and Experimental Medicine |
| spelling | doaj-art-40d37611aa6845deb5073c00bbd875ca2025-08-20T02:01:24ZengWileyAnimal Models and Experimental Medicine2576-20952024-12-017694495410.1002/ame2.12512Initial pig developmental stage influences intestinal organoid growth but not cellular compositionCamille Duchesne0Gwénaëlle Randuineau1Laurence Le Normand2Véronique Romé3Samia Laraqui4Alexis Pierre Arnaud5Gaëlle Boudry6Institut Numecan, INRAE, INSERM Univ Rennes Rennes FranceInstitut Numecan, INRAE, INSERM Univ Rennes Rennes FranceInstitut Numecan, INRAE, INSERM Univ Rennes Rennes FranceInstitut Numecan, INRAE, INSERM Univ Rennes Rennes FranceInstitut Numecan, INRAE, INSERM Univ Rennes Rennes FranceInstitut Numecan, INRAE, INSERM Univ Rennes Rennes FranceInstitut Numecan, INRAE, INSERM Univ Rennes Rennes FranceAbstract Background Intestinal organoids are promising tools in the context of animal experiment reduction but a thorough characterization of the impact of the origin of intestinal stem cells (ISC) on organoid phenotype is needed to routinely use this cellular model. Our objective was to evaluate the effect of ISC donor age on the growth, morphology and cellular composition of intestinal organoids derived from pig. Methods Organoids were derived from jejunal and colonic ISC obtained from 1‐, 7‐, 28‐, 36‐ and 180‐day‐old pigs and passaged three times. Results We first confirmed by qPCR that the expression of 18% of the >80 studied genes related to various intestinal functions differed between jejunal and colonic organoids after two passages (p < 0.05). Growth and morphology of organoids depended on intestinal location (greater number and larger organoids derived from colonic than jejunal ISC, p < 0.05) but also pig age. Indeed, when ISC were derived from young piglets, the ratio of organoids to spheroids was greater (p < 0.05), spheroids were larger during the primary culture but smaller after two passages (p < 0.05) and organoids were smaller after one passage (p > 0.05) compared to ISC from older pigs. Finally, no difference in cellular composition, evaluated by immunostaining of markers of the major intestinal cell types (absorptive, enteroendocrine and goblet cells) was observed between organoids originating from 7‐ or 180‐day‐old pigs, but differences between intestinal site origins were noticed. Conclusion In conclusion, while the age of the tissue donor affected organoid growth and morphology, it did not influence the phenotype.https://doi.org/10.1002/ame2.12512colonoiddevelopmententeroidorganoid phenotypepiglet |
| spellingShingle | Camille Duchesne Gwénaëlle Randuineau Laurence Le Normand Véronique Romé Samia Laraqui Alexis Pierre Arnaud Gaëlle Boudry Initial pig developmental stage influences intestinal organoid growth but not cellular composition Animal Models and Experimental Medicine colonoid development enteroid organoid phenotype piglet |
| title | Initial pig developmental stage influences intestinal organoid growth but not cellular composition |
| title_full | Initial pig developmental stage influences intestinal organoid growth but not cellular composition |
| title_fullStr | Initial pig developmental stage influences intestinal organoid growth but not cellular composition |
| title_full_unstemmed | Initial pig developmental stage influences intestinal organoid growth but not cellular composition |
| title_short | Initial pig developmental stage influences intestinal organoid growth but not cellular composition |
| title_sort | initial pig developmental stage influences intestinal organoid growth but not cellular composition |
| topic | colonoid development enteroid organoid phenotype piglet |
| url | https://doi.org/10.1002/ame2.12512 |
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