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 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.

塑造组织结构和功能的细胞多样性受多种微环境信号的调控。然而，在人肠道类器官中维持细胞多样性一直具有挑战性。基于天然干细胞环境中存在的微环境配体，我们为肠上皮建立了一种改良的类器官培养条件，该条件使人肠道类器官能够同时进行多向分化和自我更新。高通量筛选显示，胰岛素样生长因子1（IGF - 1）和成纤维细胞生长因子2（FGF - 2）的组合可提高人肠道干细胞的克隆形成能力和CRISPR基因组工程效率。该组合同样能够对一系列肠道类器官进行长期培养，包括大鼠小肠类器官。基于液滴的单细胞RNA测序进一步表明，在改良条件下培养的人小肠类器官中，天然细胞多样性得以保留。改良的培养方案优于传统方法，为在体内相关环境下模拟人肠道组织和疾病提供了一种可行的策略。