诱导多能干细胞（iPSC）已成为有前景的体外工具，为疾病建模和促进药物筛选提供了强大的系统。人类 iPSC 已成功分化为肺细胞和三维肺球体或类器官。肺是一个多细胞的复杂器官，在微环境的交响影响下发育。在这里，我们假设在受控微环境（cmO）（氧气和压力）中生成肺类器官会产生具有类似于肺泡的结构复杂性的多细胞类器官。 iPSC 在氧气和压力控制的微环境中按照逐步方案分化为成熟的肺类器官。在受控微环境中发育的类器官显示出复杂的肺泡结构，并进行 SFTPC、PDPN 和 KRT5 染色，表明存在肺泡上皮 II 型和 I 型细胞以及基底细胞。此外，cmO 中的基因和蛋白质表达水平也有所增加。此外，蛋白质组学的通路分析揭示了与正常培养条件下生长的相比，cmO 中肺发育特异性通路的上调。总之，通过使用受控的微环境，我们建立了一种源自 iPSC 的复杂多细胞肺类器官，作为研究肺部健康和疾病中肺泡生物学的新型细胞模型。© 2024。作者。

Induced pluripotent stem cells (iPSCs) have emerged as promising in vitro tools, providing a robust system for disease modelling and facilitating drug screening. Human iPSCs have been successfully differentiated into lung cells and three-dimensional lung spheroids or organoids. The lung is a multicellular complex organ that develops under the symphonic influence of the microenvironment. Here, we hypothesize that the generation of lung organoids in a controlled microenvironment (cmO) (oxygen and pressure) yields multicellular organoids with architectural complexity resembling the lung alveoli. iPSCs were differentiated into mature lung organoids following a stepwise protocol in an oxygen and pressure-controlled microenvironment. The organoids developed in the controlled microenvironment displayed complex alveolar architecture and stained for SFTPC, PDPN, and KRT5, indicating the presence of alveolar epithelial type II and type I cells, as well as basal cells. Moreover, gene and protein expression levels were also increased in the cmO. Furthermore, pathway analysis of proteomics revealed upregulation of lung development-specific pathways in the cmO compared to those growing in normal culture conditions. In summary, by using a controlled microenvironment, we established a complex multicellular lung organoid derived from iPSCs as a novel cellular model to study lung alveolar biology in both lung health and disease.© 2024. The Author(s).