肺泡上皮细胞易受损伤，这可能导致上皮增生、细胞凋亡和慢性炎症。在本研究中，我们开发了人类诱导多能干细胞(hiPS)细胞衍生的肺泡上皮细胞（iAECs）和基于iAECs的器官体（AOs），以测试化学物质暴露后的AEC毒性。 hiPS细胞在分化培养基中分别培养14天，并在另外14天的时间内维持iAECs基于AOs的培养状态。AOs表达了SFTPC和AQP5，并增加了SOX9、NKX2.1、GATA6、HOPX和ID2的mRNA水平。AOs暴露于9种化学物质中24小时，并使用MTT测定得出这9种化学物质的IC50值。当使用皮尔逊相关系数r值计算iAECs 2D培养和AOs 3D培养之间的相关性时，发现在3D培养中导致显著细胞存活率下降的9种化学物质在2D培养中高度相关。然后研究了AO与单核细胞的细胞毒性和一氧化氮释放。当AO与单核细胞共同暴露于铬酸钠24小时时，IC50值和一氧化氮产生量比AO单独暴露高。综上所述，基于AO的3D培养系统为了解AEC的生物学特性和建模化学物质暴露提供了有用的平台。版权所有©2023 Elsevier Ltd。

Alveolar epithelial cells (AECs) are vulnerable to injury, which can result in epithelial hyperplasia, apoptosis, and chronic inflammation. In this study, we developed human induced pluripotent stem cell (hiPS) cell-derived AECs (iAECs) and the iAECs based organoids (AOs) for testing AEC toxicity after chemical exposure. HiPS cells were cultured for 14 days with differentiation medium corresponding to each step, and the iAECs-based AOs were maintained for another 14 days. SFTPC and AQP5 were expressed in the AOs, and mRNA levels of SOX9, NKX2.1, GATA6, HOPX, and ID2 were increased. The AOs were exposed for 24 h to nine chemical substances, and IC50 values of the nine chemicals were determined using MTT assay. When the correlations between iAECs 2D culture and AOs 3D culture were calculated using Pearson's correlation coefficient r value, the nine chemicals that caused a significant decrease of cell viability in 3D culture were found to be highly correlated in 2D culture. The cytotoxicity and nitric oxide release in AO cultured with macrophages were then investigated. When AOs with macrophages were exposed to sodium chromate for 24 h, the IC50 value and nitric oxide production were higher than when the AOs were exposed alone. Taken together, the AO-based 3D culture system provides a useful platform for understanding biological characteristics of AECs and modeling chemical exposures.Copyright © 2023. Published by Elsevier Ltd.