Cell Stem Cell
Volume 23, Issue 6, 6 December 2018, Pages 787-793.e6
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Brief Report
Human Intestinal Organoids Maintain Self-Renewal Capacity and Cellular Diversity in Niche-Inspired Culture Condition

https://doi.org/10.1016/j.stem.2018.11.016Get rights and content
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Highlights

  • 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

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.

Keywords

p38 signaling
LGR5
intestinal stem cells
Paneth cells
enteroendocrine cells
tuft cells
M cells
ulcerative colitis
CRISPR-Cas9
diffusion pseudotime

Cited by (0)

5

These authors contributed equally

6

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