Human Intestinal Organoids Maintain Self-Renewal Capacity and Cellular Diversity in Niche-Inspired Culture Condition

Cellular diversity that shapes tissue architecture and function is governed by multiple niche signals. Nonetheless, maintaining cellular diversity in human intestinal organoids has been challenging. Based on niche ligands present in the natural stem cell milieu, we establish a refined organoid cultu...

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Bibliographic Details
Published in:Cell stem cell 2018-12, Vol.23 (6), p.787-793.e6
Main Authors: Fujii, Masayuki, Matano, Mami, Toshimitsu, Kohta, Takano, Ai, Mikami, Yohei, Nishikori, Shingo, Sugimoto, Shinya, Sato, Toshiro
Format: Article
Language:English
Subjects:
CRISPR-Cas9
diffusion pseudotime
enteroendocrine cells
intestinal stem cells
LGR5
M cells
p38 signaling
Paneth cells
tuft cells
ulcerative colitis
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Summary:Cellular diversity that shapes tissue architecture and function is governed by multiple niche signals. Nonetheless, maintaining cellular diversity in human intestinal organoids has been challenging. Based on niche ligands present in the natural stem cell milieu, we establish a refined organoid culture condition for intestinal epithelia that allows human intestinal organoids to concurrently undergo multi-differentiation and self-renewal. High-throughput screening reveals that the combination of insulin-like growth factor 1 (IGF-1) and fibroblast growth factor 2 (FGF-2) enhances the clonogenic capacity and CRISPR-genome engineering efficiency of human intestinal stem cells. The combination equally enables long-term culture of a range of intestinal organoids, including rat small intestinal organoids. Droplet-based single-cell RNA sequencing further illustrates the conservation of the native cellular diversity in human small intestinal organoids cultured with the refined condition. The modified culture protocol outperforms the conventional method and offers a viable strategy for modeling human intestinal tissues and diseases in an in vivo relevant context. [Display omitted] •IGF-1 and FGF-2 improve human intestinal organoid plating and genome editing efficiencies•Organoids maintain self-renewal and multi-differentiation capacity in refined condition•Refined condition enables long-term culture of healthy and diseased intestinal organoids•Human small intestinal crypts and organoids compared with droplet-based scRNA-seq Sato and colleagues develop a modified culture condition for human intestinal organoids that improves the culture efficiency and maintains their long-term multi-differentiation capacity. scRNA-seq of human small intestinal crypts and organoids demonstrates that in vivo cellular diversity can be preserved in organoids cultured with the refined condition.
ISSN:1934-5909
1875-9777
DOI:10.1016/j.stem.2018.11.016