Department of Respiratory Medicine, Graduate School of Medicine, Kyoto University, Kyoto, Japan; Department of Drug Discovery for Lung Diseases, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Department of Respiratory Medicine, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Methods Cell Biol. 2020;159:115-141. doi: 10.1016/bs.mcb.2020.02.004. Epub 2020 Mar 17.
The lung consists of branched structures that are anatomically, developmentally and functionally divided into airway and alveolar regions. Each region contributes to lung-specific functions involving a defense system and gas exchange, and their dysfunction can cause fatal lung diseases. In the alveolar region, the cuboidal alveolar type 2 (AT2) cells account for 90% of the alveolar epithelial cells and serve as the tissue stem cells secreting pulmonary surfactant, and flattened alveolar type I (AT1) cells cover most of the alveolar surface directly contributing to gas exchange adjacent to capillary vessels. It has been difficult to culture alveolar epithelial cells in vitro, as the lineage-specific features of those cells are rapidly lost in a conventional two-dimensional culture setting. The culture of alveolar organoids (AOs) is an emerging technique that can help maintain the features of alveolar epithelial cells in vitro, and their application to human disease modeling is eagerly awaited. We herein describe our method of generating and culturing alveolar epithelial cells and AOs derived from human induced pluripotent stem cells (iPSCs). iPSCs derived from lung disease patients, including those with rare genetic diseases, will make help elucidate the disease mechanism and hopefully identify therapeutic targets.
肺由分支结构组成，这些结构在解剖学、发育和功能上分为气道和肺泡区域。每个区域都有助于涉及防御系统和气体交换的肺特异性功能，其功能障碍可导致致命的肺部疾病。在肺泡区域，立方肺泡 II 型 (AT2) 细胞占肺泡上皮细胞的 90%，作为分泌肺表面活性剂的组织干细胞，扁平肺泡 I 型 (AT1) 细胞覆盖大部分肺泡表面，直接与毛细血管相邻促进气体交换。由于这些细胞的谱系特异性特征在传统的二维培养环境中迅速丧失，因此很难在体外培养肺泡上皮细胞。肺泡类器官 (AO) 的培养是一种新兴技术，可以帮助维持体外肺泡上皮细胞的特征，人们急切地期待其在人类疾病建模中的应用。我们在此描述了从人诱导多能干细胞 (iPSC) 生成和培养肺泡上皮细胞和 AO 的方法。来自肺部疾病患者（包括罕见遗传疾病患者）的 iPSC 将有助于阐明疾病机制，并有望确定治疗靶点。
