| Autor: |
Ament AL; Universities of Giessen and Marburg Lung Center, Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Cardio-Pulmonary Institute (CPI) , Giessen, Hessen, Germany.; German Center for Lung Research, Institute for Lung Health (ILH), Giessen, Hessen, Germany.; Rheinische Friedrich-Wilhelms-Universität Bonn LIMES, Organoid Biology, Bonn, Germany., Heiner M; Universities of Giessen and Marburg Lung Center, Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Giessen, Hessen, Germany., Hessler MC; Universities of Giessen and Marburg Lung Center, Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Giessen, Hessen, Germany., Alexopoulos I; Universities of Giessen and Marburg Lung Center, Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Giessen, Hessen, Germany.; German Center for Lung Research, Institute for Lung Health (ILH), Giessen, Hessen, Germany., Steeg K; Universities of Giessen and Marburg Lung Center, Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Giessen, Hessen, Germany., Gärtner U; Justus Liebig Universitat Giessen, Institute for Anatomy and Cell Biology, German Center for Lung Research, Excellence Cluster Cardio-Pulmonary Institute (CPI), Giessen, Germany., Vazquez-Armendariz AI; Rheinische Friedrich-Wilhelms-Universität Bonn LIMES, Organoid Biology, Bonn, Germany.; Universities of Giessen and Marburg Lung Center, Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Cardio-Pulmonary Institute (CPI) , Giessen, Hessen, Germany.; German Center for Lung Research, Institute for Lung Health (ILH), Giessen, Hessen, Germany; vazquez@uni-bonn.de., Herold S; Universities of Giessen and Marburg Lung Center, Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Cardio-Pulmonary Institute (CPI) , Giessen, Hessen, Germany.; German Center for Lung Research, Institute for Lung Health (ILH), Giessen, Hessen, Germany. |
| Abstrakt: |
Organoid 3D systems are powerful platforms to study development and disease. Recently, the complexity of lung organoid models derived from adult mouse and human stem cells has increased substantially in terms of cellular composition and structural complexity. However, a murine lung organoid system with a clear integrated endothelial compartment is still missing. Here, we describe a novel method that adds another level of intricacy to our published bronchioalveolar lung organoid (BALO) model by microinjection of FACS-sorted lung endothelial cells (ECs) into differentiated organoid cultures. Before microinjection, ECs obtained from the lung homogenate (LH) of young mice expressed typical ECs markers such as CD31 and vascular endothelial (VE)-Cadherin and showed tube formation capacity. Following microinjection, ECs surrounded BALO´s alveolar-like compartment aligning with both alveolar epithelial cells type I (AECI) and type II (AECII), as demonstrated by confocal and electron microscopy. Notably, expression of Car4 and Aplnr was as well detected, suggesting presence of EC microvascular phenotypes in the cultured ECs. Moreover, upon epithelial cell injury by lipopolysaccharides (LPS) and influenza A virus (IV), endothelialized BALO (eBALO) released proinflammatory cytokines leading to the upregulation of the intercellular adhesion molecule 1 (ICAM-1) in ECs. In summary, we characterized for the first time a organoid model that incorporates ECs into the alveolar structures of lung organoids, not only increasing our previous model ́s cellular and structural complexity but also providing a suitable niche to model lung endothelium responses to injury ex vivo. |
