Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System
Intestinal organoids offer great promise for disease-modelling-based host–pathogen interactions and nutritional research for feed efficiency measurement in livestock and regenerative medicine for therapeutic purposes. However, very limited studies are available on the functional characterisation and...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-07-01
|
| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/11/7/2115 |
| _version_ | 1827688110219067392 |
|---|---|
| author | Bo-Ram Lee Hyeon Yang Sang-In Lee Inamul Haq Sun-A Ock Hayeon Wi Hwi-Cheul Lee Poongyeon Lee Jae-Gyu Yoo |
| author_facet | Bo-Ram Lee Hyeon Yang Sang-In Lee Inamul Haq Sun-A Ock Hayeon Wi Hwi-Cheul Lee Poongyeon Lee Jae-Gyu Yoo |
| author_sort | Bo-Ram Lee |
| collection | DOAJ |
| description | Intestinal organoids offer great promise for disease-modelling-based host–pathogen interactions and nutritional research for feed efficiency measurement in livestock and regenerative medicine for therapeutic purposes. However, very limited studies are available on the functional characterisation and three-dimensional (3D) expansion of adult stem cells in livestock species compared to other species. Intestinal crypts derived from intestinal organoids under a 3D culture system from the small intestine in adult bovine were successfully established and characterised for functionality testing, including the cellular potentials and genetic properties based on immunohistochemistry, immunocytochemistry, epithelial barrier permeability assay, QuantSeq 3′ mRNA-Seq. data and quantitative reverse transcription-polymerase chain reaction. Intestinal organoids were long-term cultivated over several passages of culture without loss of the recapitulating capacity of crypts, and they had the specific expression of several specific markers involved in intestinal stem cells, intestinal epithelium, and nutrient absorption. In addition, they showed the key functionality with regard to a high permeability for compounds of up to FITC-dextran 4 kDa, while FITC-dextran 40 kDa failed to enter the organoid lumen and revealed that the genetic properties of bovine intestinal organoids were highly similar to those of in vivo. Collectively, these results provide a reliable method for efficient isolation of intestinal crypts from the small intestine and robust 3D expansion of intestinal organoids in adult bovine and demonstrate the in vitro 3D organoids mimics the in vivo tissue topology and functionality. Finally, intestinal organoids are potential alternatives to in vivo systems and will be facilitated as the practical model to replace animal experiments for various purposes in the fields of animal biotechnology. |
| first_indexed | 2024-03-10T09:48:20Z |
| format | Article |
| id | doaj.art-cfa5f45c432a41b7bdfec389913e2549 |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T09:48:20Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-cfa5f45c432a41b7bdfec389913e25492023-11-22T03:02:39ZengMDPI AGAnimals2076-26152021-07-01117211510.3390/ani11072115Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo SystemBo-Ram Lee0Hyeon Yang1Sang-In Lee2Inamul Haq3Sun-A Ock4Hayeon Wi5Hwi-Cheul Lee6Poongyeon Lee7Jae-Gyu Yoo8Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaDepartment of Animal Biotechnology, Kyungpook National University, Sangju-si 37224, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaPlanning and Coordination Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaAnimal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju-gun 55365, KoreaIntestinal organoids offer great promise for disease-modelling-based host–pathogen interactions and nutritional research for feed efficiency measurement in livestock and regenerative medicine for therapeutic purposes. However, very limited studies are available on the functional characterisation and three-dimensional (3D) expansion of adult stem cells in livestock species compared to other species. Intestinal crypts derived from intestinal organoids under a 3D culture system from the small intestine in adult bovine were successfully established and characterised for functionality testing, including the cellular potentials and genetic properties based on immunohistochemistry, immunocytochemistry, epithelial barrier permeability assay, QuantSeq 3′ mRNA-Seq. data and quantitative reverse transcription-polymerase chain reaction. Intestinal organoids were long-term cultivated over several passages of culture without loss of the recapitulating capacity of crypts, and they had the specific expression of several specific markers involved in intestinal stem cells, intestinal epithelium, and nutrient absorption. In addition, they showed the key functionality with regard to a high permeability for compounds of up to FITC-dextran 4 kDa, while FITC-dextran 40 kDa failed to enter the organoid lumen and revealed that the genetic properties of bovine intestinal organoids were highly similar to those of in vivo. Collectively, these results provide a reliable method for efficient isolation of intestinal crypts from the small intestine and robust 3D expansion of intestinal organoids in adult bovine and demonstrate the in vitro 3D organoids mimics the in vivo tissue topology and functionality. Finally, intestinal organoids are potential alternatives to in vivo systems and will be facilitated as the practical model to replace animal experiments for various purposes in the fields of animal biotechnology.https://www.mdpi.com/2076-2615/11/7/2115bovineintestinal stem cellsorganoidcharacterisationgene expression |
| spellingShingle | Bo-Ram Lee Hyeon Yang Sang-In Lee Inamul Haq Sun-A Ock Hayeon Wi Hwi-Cheul Lee Poongyeon Lee Jae-Gyu Yoo Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System Animals bovine intestinal stem cells organoid characterisation gene expression |
| title | Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System |
| title_full | Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System |
| title_fullStr | Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System |
| title_full_unstemmed | Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System |
| title_short | Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System |
| title_sort | robust three dimensional 3d expansion of bovine intestinal organoids an in vitro model as a potential alternative to an in vivo system |
| topic | bovine intestinal stem cells organoid characterisation gene expression |
| url | https://www.mdpi.com/2076-2615/11/7/2115 |
| work_keys_str_mv | AT boramlee robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT hyeonyang robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT sanginlee robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT inamulhaq robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT sunaock robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT hayeonwi robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT hwicheullee robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT poongyeonlee robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem AT jaegyuyoo robustthreedimensional3dexpansionofbovineintestinalorganoidsaninvitromodelasapotentialalternativetoaninvivosystem |