McVicar Rachael N, Smith Emily, Melameka Melina, Bush Anne, Goetz Grace, Constantino Gailan, Kumar Matangi, Kwong Elizabeth, Snyder Evan Y, Leibel Sandra L
Sanford Consortium for Regenerative Medicine; Sanford Burnham Prebys Medical Discovery Institute.
Sanford Consortium for Regenerative Medicine; Sanford Burnham Prebys Medical Discovery Institute;
J Vis Exp. 2024 Dec 6(214). doi: 10.3791/67247.
Human lung tissue is composed of an interconnected network of epithelium, mesenchyme, endothelium, and immune cells from the upper airway of the nasopharynx to the smallest alveolar sac. Interactions between these cells are crucial in lung development and disease, acting as a barrier against harmful chemicals and pathogens. Current in vitro co-culture models utilize immortalized cell lines with different biological backgrounds, which may not accurately represent the cellular milieu or interactions of the lung. We differentiated human iPSCs into 3D lung organoids (containing both epithelium and mesenchyme), endothelial cells, and macrophages. These were co-cultured in an air-liquid interface (ALI) format to form an epithelial/mesenchymal apical barrier invested with macrophages and a basolateral endothelial barrier (iAirway). iPSC-derived iAirways showed a reduction in barrier integrity in response to infection with respiratory viruses and cigarette toxins. This multi-lineage lung co-culture system provides a platform for studying cellular interactions, signaling pathways, and molecular mechanisms underlying lung development, homeostasis, and disease progression. iAirways closely mimic human physiology and cellular interactions, can be generated from patient-derived iPSC's, and can be customized to include different cell types of the airway. Overall, iPSC-derived iAirway models offer a versatile and powerful tool for studying barrier integrity to better understand genetic drivers for disease, pathogen response, immune regulation, and drug discovery or repurposing in vitro, with the potential to advance our understanding and treatment of airway diseases.
人类肺组织由从鼻咽部上呼吸道到最小肺泡囊的上皮细胞、间充质细胞、内皮细胞和免疫细胞相互连接的网络组成。这些细胞之间的相互作用在肺发育和疾病中至关重要，可作为抵御有害化学物质和病原体的屏障。当前的体外共培养模型使用具有不同生物学背景的永生化细胞系，这可能无法准确代表肺的细胞环境或细胞间相互作用。我们将人类诱导多能干细胞分化为三维肺类器官（包含上皮细胞和间充质细胞）、内皮细胞和巨噬细胞。将这些细胞以气液界面（ALI）形式进行共培养，形成一个由巨噬细胞构成的上皮/间充质顶端屏障和一个基底侧内皮屏障（iAirway）。源自诱导多能干细胞的iAirway在受到呼吸道病毒感染和香烟毒素刺激时，屏障完整性降低。这种多谱系肺共培养系统为研究肺发育、稳态和疾病进展背后的细胞相互作用、信号通路及分子机制提供了一个平台。iAirway紧密模拟人类生理学和细胞间相互作用，可从患者来源的诱导多能干细胞生成，还可定制以纳入气道的不同细胞类型。总体而言，源自诱导多能干细胞的iAirway模型为研究屏障完整性提供了一种通用且强大的工具，有助于更好地理解疾病的遗传驱动因素、病原体反应、免疫调节以及体外药物发现或重新利用，有潜力增进我们对气道疾病的理解和治疗。
