Human organoid model systems lack important cell types that, in the embryo, are incorporated into organ tissues during development. We developed an organoid assembly approach starting with cells from the three primary germ layers—enteric neuroglial, mesenchymal, and epithelial precursors—that were derived separately from human pluripotent stem cells (PSCs). From these three cell types, we generated human antral and fundic gastric tissue containing differentiated glands surrounded by layers of smooth muscle containing functional enteric neurons that controlled contractions of the engineered antral tissue. Using this experimental system, we show that human enteric neural crest cells (ENCCs) promote mesenchyme development and glandular morphogenesis of antral stomach organoids. Moreover, ENCCs can act directly on the foregut to promote a posterior fate, resulting in organoids with a Brunner’s gland phenotype. Thus, germ layer components that are derived separately from PSCs can be used for tissue engineering to generate complex human organoids.

人体类器官模型系统缺乏重要的细胞类型，在胚胎中，这些细胞类型在发育过程中被纳入器官组织。我们开发了一种类器官组装方法，从三个主要胚层（肠神经胶质细胞、间充质细胞和上皮前体细胞）的细胞开始，这些细胞分别来源于人类多能干细胞 (PSC)。从这三种细胞类型中，我们生成了人类胃窦和胃底胃组织，其中包含分化的腺体，这些腺体被平滑肌层包围，平滑肌层含有控制工程胃窦组织收缩的功能性肠神经元。使用该实验系统，我们表明人肠神经嵴细胞 (ENCC) 促进胃窦类器官的间充质发育和腺体形态发生。而且，ENCC 可以直接作用于前肠以促进后部命运，从而产生具有 Brunner 腺体表型的类器官。因此，从 PSC 中分离出来的胚层成分可用于组织工程以生成复杂的人体类器官。