Chemically-defined and scalable culture system for intestinal stem cells derived from human intestinal organoids

• Published in: Nature communications Vol. 15; no. 1; pp. 799 - 14
• Main Authors: Kwon, Ohman, Lee, Hana, Jung, Jaeeun, Son, Ye Seul, Jeon, Sojeong, Yoo, Won Dong, Son, Naeun, Jung, Kwang Bo, Choi, Eunho, Lee, In-Chul, Kwon, Hyung-Jun, Kim, Chuna, Lee, Mi-Ok, Cho, Hyun-Soo, Kim, Dae Soo, Son, Mi-Young
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 27-01-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 13/100; 14/1; 14/35; 38/109; 38/23; 38/39; 38/77; 38/90; 38/91; 42/100; 42/109; 631/136/532/2437; 631/1647/767/2199; 631/532/2437; 631/61/2320; 64/60; 82/51; 82/80; Cell culture; Cell Differentiation; Cellular structure; Coronaviruses; Culture; Epithelium; Genetic modification; Genome editing; Humanities and Social Sciences; Humans; Intestinal Mucosa; Intestine; Intestines; Medical research; multidisciplinary; Organoids; Pluripotency; Pluripotent Stem Cells; Regenerative medicine; Reproducibility; Reproducibility of Results; Science; Science (multidisciplinary); Severe acute respiratory syndrome; Severe acute respiratory syndrome coronavirus 2; Small intestine transplantation; Stem cells; Villus; Viral diseases
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-45103-7

Three-dimensional human intestinal organoids (hIO) are widely used as a platform for biological and biomedical research. However, reproducibility and challenges for large-scale expansion limit their applicability. Here, we establish a human intestinal stem cell (ISC) culture method expanded under feeder-free and fully defined conditions through selective enrichment of ISC populations (ISC
3D-hIO
) within hIO derived from human pluripotent stem cells. The intrinsic self-organisation property of ISC
3D-hIO
, combined with air-liquid interface culture in a minimally defined medium, forces ISC
3D-hIO
to differentiate into the intestinal epithelium with cellular diversity, villus-like structure, and barrier integrity. Notably, ISC
3D-hIO
is an ideal cell source for gene editing to study ISC biology and transplantation for intestinal diseases. We demonstrate the intestinal epithelium differentiated from ISC
3D-hIO
as a model system to study severe acute respiratory syndrome coronavirus 2 viral infection. ISC
3D-hIO
culture technology provides a biological tool for use in regenerative medicine and disease modelling.
Challenges in reproducibility and large-scale expansion limit the current applicability of human intestinal organoids. Here, the authors present a feeder-free, chemically-defined culture method for enrichment of intestinal stem cells isolated from 3D human intestinal organoids.