Organoids and organs-on-a-chip have emerged as powerful tools for modeling human gut physiology and disease in vitro. Although physiologically relevant, these systems often lack the environmental milieu, spatial organization, cell type diversity, and maturity necessary for mimicking human intestinal mucosa. To instead generate models closely resembling in vivo tissue, we herein integrated organoid and organ-on-a-chip technology to develop an advanced human organoid model, called “mini-colons.” By employing an asymmetric stimulation with growth factors, we greatly enhanced tissue longevity and replicated in vivo-like diversity and patterning of proliferative and differentiated cell types. Mini-colons contain abundant mucus-producing goblet cells and, signifying mini-colon maturation, single-cell RNA sequencing reveals emerging mature and functional colonocytes. This methodology is expanded to generate microtissues from the small intestine and incorporate additional microenvironmental components. Finally, our bioengineered organoids provide a precise platform to systematically study human gut physiology and pathology, and a reliable preclinical model for drug safety assessment.

类器官和芯片器官已成为体外模拟人类肠道生理学和疾病的强大工具。尽管具有生理相关性，但这些系统通常缺乏模仿人类肠粘膜所需的环境环境、空间组织、细胞类型多样性和成熟度。为了生成与体内组织非常相似的模型，我们整合了类器官和芯片上器官技术，开发了一种先进的人类类器官模型，称为“迷你结肠”。通过使用生长因子的不对称刺激，我们极大地延长了组织寿命，并复制了类似体内的增殖和分化细胞类型的多样性和模式。小结肠含有丰富的产生粘液的杯状细胞，标志着小结肠的成熟，单细胞 RNA 测序揭示了正在出现的成熟且有功能的结肠细胞。该方法已扩展到从小肠生成微组织并纳入其他微环境成分。最后，我们的生物工程类器官为系统研究人类肠道生理学和病理学提供了精确的平台，并为药物安全性评估提供了可靠的临床前模型。