Earticle

초록

영어

Recent progress has been made to establish intestinal organoids for an in vitro model as a potential alternative to an in vivo system in animals. We previously reported a reliable method for the isolation of intestinal crypts from the small intestine and robust three-dimensional (3D) expansion of intestinal organoids (basal-out) in adult bovines. The present study aimed to establish next-generation intestinal organoids for practical applications in disease modeling-based host-pathogen interactions and feed efficiency measurements. In this study, we developed a rapid and convenient method for the efficient generation of intestinal organoids through the modulation of the Wnt signaling pathway and continuous apical-out intestinal organoids. Remarkably, the intestinal epithelium only takes 3-4 days to undergo CHIR (1 µM) treatment as a Wnt activator, which is much shorter than that required for spontaneous differentiation (7 days). Subsequently, we successfully established an apical-out bovine intestinal organoid culture system through suspension culture without Matrigel matrix, indicating an apical-out membrane on the surface. Collectively, these results demonstrate the efficient generation and next-generation of bovine intestinal organoids and will facilitate their potential use for various purposes, such as disease modeling, in the field of animal biotechnology.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Maintenance and cultivation of bovine intestinal organoids
Morphological classification of bovine intestinal organoids and CHIR treatment
RNA isolation
Quantitative RT-PCR
Generation of apical-out intestinal organoids
Epithelial barrier permeability assay using FITC-dextran
Statistical analysis
RESULTS AND DISCUSSION
Morphological classification of bovine intestinal organoids
Effect on Wnt activator treatment in bovine intestinal organoids
Gene expression profiling and generation of apical-out intestinal organoids
CONCLUSION
SUPPLEMENTARY MATERIALS
REFERENCES

키워드

저자

• Kang Won Park [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration ]
• Hyeon Yang [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration ]
• Hayeon Wi [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration ]
• Sun A Ock [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration ]
• Poongyeon Lee [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration ]
• In-Sul Hwang [ Columbia Center for Translational Immunology, Columbia University Irving Medical Center, Columbia University, New York 10032, USA ]
• Bo Ram Lee [ Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration ] Corresponding Author

참고문헌

발행기관

간행물

관심저널 등록

• 간행물명

  Journal of Animal Reproduction and Biotechnology

• 간기

  계간

• pISSN

  2671-4639

• eISSN

  2671-4663

• 수록기간

  2019~2025

• 십진분류

  KDC 527 DDC 636