Endothelialized Bronchioalveolar Lung Organoids Model Endothelial Cell Responses to Injury
Abstract
Organoid three-dimensional 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 of young mice expressed typical EC markers such as CD31 and vascular endothelial cadherin and showed tube formation capacity. Following microinjection, ECs surrounded the BALO’s alveolar-like compartment, aligning with type I and type II alveolar epithelial cells, as demonstrated by confocal and electron microscopy. Notably, expression of Car4 and Aplnr was as well detected, suggesting the presence of EC microvascular phenotypes in the cultured ECs. Moreover, upon epithelial cell injury by LPS and influenza A virus, endothelialized BALOs released proinflammatory cytokines, leading to the upregulation ICAM-1 (intercellular adhesion molecule 1) in ECs. In summary, we characterized for the first time an 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.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Endothelial cells have been used to endothelialize different organoids.
“Endothelial cells have been used to endothelialize different organoids.[ 67 , 68 ]”
From Coordinated Development of Immune Cell Populations in Vascularized Skin Organoids from Human Induced Pluripotent Stem Cells (2025) · cites this paper as [68]
Cites
Reconstituting organ-level lung functions on a chip
Huh, D., Matthews, B. D., Mammoto, A., Montoya-Zavala, M., Hsin, H. Y., & Ingber, D. E. (2010) · Science · doi:10.1126/science.1188302 · Cited by 4,250 · see citing works
Trends in Care Practices, Morbidity, and Mortality of Extremely Preterm Neonates, 1993-2012
Stoll, B. J., Hansen, N. I., Bell, E. F., Walsh, M. C., Carlo, W. A., Shankaran, S., Laptook, A. R., Sánchez, P. J., Van Meurs, K. P., Wyckoff, M., et al. (2015) · JAMA · doi:10.1001/jama.2015.10244 · Cited by 2,791 · see citing works
Human organoids: model systems for human biology and medicine
Kim, J., Koo, B.-K., & Knoblich, J. A. (2020) · Nature Reviews Molecular Cell Biology · doi:10.1038/s41580-020-0259-3 · Cited by 2,187 · see citing works
Evolving functions of endothelial cells in inflammation
Pober, J. S., & Sessa, W. C. (2007) · Nature Reviews Immunology · doi:10.1038/nri2171 · Cited by 2,022 · see citing works
Zhao, Z., Chen, X., Dowbaj, A. M., Sljukic, A., Bratlie, K., Lin, L., Fong, E. L. S., Balachander, G. M., Chen, Z., Soragni, A., et al. (2022) · Nature Reviews Methods Primers · doi:10.1038/s43586-022-00174-y · Cited by 1,001 · see citing works
Bioprinting 3D microfibrous scaffolds for engineering endothelialized myocardium and heart-on-a-chip
Zhang, Y. S., Arneri, A., Bersini, S., Shin, S.-R., Zhu, K., Goli-Malekabadi, Z., Aleman, J., Colosi, C., Busignani, F., Dell’Erba, V., et al. (2016) · Biomaterials · doi:10.1016/j.biomaterials.2016.09.003 · Cited by 957 · see citing works
Hogan, B. L. M., Barkauskas, C. E., Chapman, H. A., Epstein, J. A., Jain, R., Hsia, C. C. W., Niklason, L., Calle, E., Le, A., Randell, S. H., et al. (2014) · Cell Stem Cell · doi:10.1016/j.stem.2014.07.012 · Cited by 876 · see citing works
Michiels, C. (2003) · Journal of Cellular Physiology · doi:10.1002/jcp.10333 · Cited by 774 · see citing works
Teijaro, J. R., Walsh, K. B., Cahalan, S., Fremgen, D. M., Roberts, E., Scott, F., Martinborough, E., Peach, R., Oldstone, M. B. A., & Rosen, H. (2011) · Cell · doi:10.1016/j.cell.2011.08.015 · Cited by 688 · see citing works
Immunomodulation by endothelial cells — partnering up with the immune system?
Amersfoort, J., Eelen, G., & Carmeliet, P. (2022) · Nature reviews. Immunology · doi:10.1038/s41577-022-00694-4 · Cited by 549 · see citing works
Lee, J., Bhang, D. H., Beede, A. M., Huang, T. L., Stripp, B. R., Bloch, K. D., Wagers, A. J., Tseng, Y., Ryeom, S. W., & Kim, C. F. (2014) · Cell · doi:10.1016/j.cell.2013.12.039 · Cited by 491 · see citing works
Capillary cell-type specialization in the alveolus
Gillich, A., Zhang, F., Farmer, C. G., Travaglini, K. J., Tan, S. Y., Gu, M., Zhou, B., Feinstein, J. A., Krasnow, M. A., & Metzger, R. J. (2020) · Nature · doi:10.1038/s41586-020-2822-7 · Cited by 490 · see citing works
Engineering Organoid Vascularization
Grebenyuk, S., & Ranga, A. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00039 · Cited by 295 · see citing works
Early Fetal Development of Lung Vasculature
deMello, D. E., Sawyer, D., Galvin, N. J., & Reid, L. (1997) · American Journal of Respiratory Cell and Molecular Biology · doi:10.1165/ajrcmb.16.5.9160839 · Cited by 293 · see citing works
Wörsdörfer, P., Dalda, N., Kern, A., Krüger, S., Wagner, N., Kwok, C. K., Henke, E., & Ergün, S. (2019) · Scientific Reports · doi:10.1038/s41598-019-52204-7 · Cited by 278 · see citing works
Parallels of craniofacial maldevelopment in down syndrome and Ts65Dn mice
Richtsmeier, J. T., Baxter, L. L., & Reeves, R. H. (2000) · Developmental Dynamics · doi:10.1002/(sici)1097-0177(200002)217:2 · Cited by 255 · see citing works
Epithelial Vegfa Specifies a Distinct Endothelial Population in the Mouse Lung
Ellis, L. V., Cain, M. P., Hutchison, V., Flodby, P., Crandall, E. D., Borok, Z., Zhou, B., Ostrin, E. J., Wythe, J. D., & Chen, J. (2020) · Developmental Cell · doi:10.1016/j.devcel.2020.01.009 · Cited by 234 · see citing works
Defining the role of pulmonary endothelial cell heterogeneity in the response to acute lung injury
Niethamer, T. K., Stabler, C. T., Leach, J. P., Zepp, J. A., Morley, M. P., Babu, A., Zhou, S., & Morrisey, E. E. (2020) · eLife · doi:10.7554/elife.53072 · Cited by 234 · see citing works
Endothelial microRNAs regulating the NF‐κB pathway and cell adhesion molecules during inflammation
Zhong, L., Simard, M. J., & Huot, J. (2018) · The FASEB Journal · doi:10.1096/fj.201701536r · Cited by 212 · see citing works
Pulmonary vascular endothelium: the orchestra conductor in respiratory diseases
Huertas, A., Guignabert, C., Barberà, J. A., Bärtsch, P., Bhattacharya, J., Bhattacharya, S., Bonsignore, M. R., Dewachter, L., Dinh‐Xuan, A. T., Dorfmüller, P., et al. (2018) · European Respiratory Journal · doi:10.1183/13993003.00745-2017 · Cited by 205 · see citing works
Leukocyte trafficking to the lungs and beyond: lessons from influenza for COVID-19
Alon, R., Sportiello, M., Kozlovski, S., Kumar, A., Reilly, E. C., Zarbock, A., Garbi, N., & Topham, D. J. (2021) · Nature Reviews Immunology · doi:10.1038/s41577-020-00470-2 · Cited by 197 · see citing works
Dye, B. R., Dedhia, P. H., Miller, A. J., Nagy, M. S., White, E. S., Shea, L. D., & Spence, J. R. (2016) · eLife · doi:10.7554/elife.19732 · Cited by 191 · see citing works
Human airway organoid engineering as a step toward lung regeneration and disease modeling
Tan, Q., Choi, K. M., Sicard, D., & Tschumperlin, D. J. (2017) · Biomaterials · doi:10.1016/j.biomaterials.2016.10.046 · Cited by 191 · see citing works
Gorina, R., Lyck, R., Vestweber, D., & Engelhardt, B. (2013) · The Journal of Immunology · doi:10.4049/jimmunol.1300858 · Cited by 182 · see citing works
Recent advances in lung organoid development and applications in disease modeling
Vazquez‐Armendariz, A. I., & Tata, P. R. (2023) · Journal of Clinical Investigation · doi:10.1172/jci170500 · Cited by 128 · see citing works
Self-citationHuman Blood Vessel Organoids Penetrate Human Cerebral Organoids and Form a Vessel-Like System
Ahn, Y.-L., An, J.-H., Yang, H.-J., Lee, D. G., Kim, J., Koh, H., Park, Y.-H., Song, B., Sim, B.-W., Lee, H. J., et al. (2021) · Cells · doi:10.3390/cells10082036 · Cited by 105 · see citing works
VEGF and endothelium-derived retinoic acid regulate lung vascular and alveolar development
Yun, E. J., Lorizio, W., Seedorf, G., Abman, S. H., & Vu, T. H. (2015) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00229.2015 · Cited by 90 · see citing works
Mußbacher, M., Schossleitner, K., Kral-Pointner, J. B., Salzmann, M., Schrammel, A., & Schmid, J. A. (2022) · Current Atherosclerosis Reports · doi:10.1007/s11883-022-01023-9 · Cited by 86 · see citing works
Vascular Niche in Lung Alveolar Development, Homeostasis, and Regeneration
Mammoto, A., & Mammoto, T. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00318 · Cited by 85 · see citing works
Role of alveolar epithelial ICAM-1 in lipopolysaccharide-induced lung inflammation
Beck‐Schimmer, B., Madjdpour, C., Kneller, S., Ziegler, U., Pasch, T., Wuthrich, R. P., Ward, P. A., & Schimmer, R. C. (2002) · European Respiratory Journal · doi:10.1183/09031936.02.00236602 · Cited by 79 · see citing works
Vazquez‐Armendariz, A. I., Heiner, M., Agha, E. E., Salwig, I., Hoek, A., Hessler, M. C., Shalashova, I., Shrestha, A., Carraro, G., Mengel, J. P., et al. (2020) · The EMBO Journal · doi:10.15252/embj.2019103476 · Cited by 77 · see citing works
Self-citationIsolation of murine lung endothelial cells
Fehrenbach, M. L., Cao, G., Williams, J. T., Finklestein, J. M., & DeLisser, H. M. (2009) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.90613.2008 · Cited by 73 · see citing works
Direct derivation of human alveolospheres for SARS-CoV-2 infection modeling and drug screening
Ebisudani, T., Sugimoto, S., Haga, K., Mitsuishi, A., Takai‐Todaka, R., Fujii, M., Toshimitsu, K., Hamamoto, J., Sugihara, K., Hishida, T., et al. (2021) · Cell Reports · doi:10.1016/j.celrep.2021.109218 · Cited by 69 · see citing works
A simple protocol for isolating mouse lung endothelial cells
Wang, J., Niu, N., Xu, S., & Jin, Z. G. (2019) · Scientific Reports · doi:10.1038/s41598-018-37130-4 · Cited by 68 · see citing works
Vasculogenesis in kidney organoids upon transplantation
Koning, M., Dumas, S. J., Avramut, M. C., Koning, R. I., Meta, E., Lievers, E., Wiersma, L. E., Borri, M., Liang, X., Xie, L. Y., et al. (2022) · npj Regenerative Medicine · doi:10.1038/s41536-022-00237-4 · Cited by 62 · see citing works
Wee, H., Oh, H., Jo, J., & Jun, C. (2009) · Experimental & Molecular Medicine · doi:10.3858/emm.2009.41.5.038 · Cited by 51 · see citing works
The Role of PGE2 in Alveolar Epithelial and Lung Microvascular Endothelial Crosstalk
Bärnthaler, T., Marić, J., Platzer, W., Kónya, V., Theiler, A., Hasenoehrl, C., Gottschalk, B., Trautmann, S., Schreiber, Y., Graier, W. F., et al. (2017) · Scientific Reports · doi:10.1038/s41598-017-08228-y · Cited by 49 · see citing works
Soluble ECM promotes organotypic formation in lung alveolar model
Valdoz, J. C., Franks, N. A., Cribbs, C. G., Jacobs, D. J., Dodson, E. L., Knight, C. J., Poulson, P. D., Garfield, S. R., Johnson, B. C., Hemeyer, B. M., et al. (2022) · Biomaterials · doi:10.1016/j.biomaterials.2022.121464 · Cited by 37 · see citing works
Human alveolar progenitors generate dual lineage bronchioalveolar organoids
Hoffmann, K., Obermayer, B., Hönzke, K., Fatykhova, D., Demir, Z., Löwa, A., Alves, L. G. T., Wyler, E., López-Rodríguez, E., Mieth, M., et al. (2022) · Communications Biology · doi:10.1038/s42003-022-03828-5 · Cited by 28 · see citing works
Rational engineering of lung alveolar epithelium
Leiby, K. L., Yuan, Y., Ng, R., Raredon, M. S. B., Adams, T. S., Baevova, P., Greaney, A. M., Hirschi, K. K., Campbell, S. G., Kaminski, N., et al. (2023) · npj Regenerative Medicine · doi:10.1038/s41536-023-00295-2 · Cited by 26 · see citing works
Development and Functional Characterization of Fetal Lung Organoids
Laube, M., Pietsch, S., Pannicke, T., Thome, U. H., & Fabian, C. (2021) · Frontiers in Medicine · doi:10.3389/fmed.2021.678438 · Cited by 20 · see citing works
Tsikis, S. T., Hirsch, T. I., Fligor, S. C., Quigley, M., & Puder, M. (2022) · Frontiers in Molecular Biosciences · doi:10.3389/fmolb.2022.1093369 · Cited by 19 · see citing works
Isolation of Endothelial Cells from Mouse Lung
Cao, G., Abraham, V., & DeLisser, H. M. (2014) · Current Protocols in Toxicology · doi:10.1002/0471140856.tx2402s61 · Cited by 7 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Immunological and regenerative properties of lung stem cells
Hao, D., Liu, Y., Li, L., Stripp, B. R., & Chen, H. (2025) · Physiological Reviews · doi:10.1152/physrev.00056.2024 · Cited by 19 · see citing works
Mostina, M., Sun, J., Sim, S. L., Ahmed, I. A., Souza‐Fonesca‐Guimaraes, F., Wolvetang, E. J., Brown, J., Kumari, S., Khosrotehrani, K., & Shafiee, A. (2025) · Advanced Healthcare Materials · doi:10.1002/adhm.202502108 · Cited by 17 · see citing works
Organoids: physiologically relevant ex vivo models for viral disease research
Wang, Y., Peng, D., Li, M., Meng, Y., Li, T., Li, S., Qiu, H., & Li, L. (2025) · Journal of Virology · doi:10.1128/jvi.01132-25 · Cited by 11 · see citing works
Endothelial specialization and angiocrine factors are critical for vascularized organoid models
Fitzsimmons, R. L., & Hudson, J. E. (2025) · Cell Reports · doi:10.1016/j.celrep.2025.116410 · Cited by 9 · see citing works
Budeus, B., Kroepel, C., Sevindik, Z. F., Buttler, L. F., & Klein, D. (2025) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2025.1684315 · Cited by 6 · see citing works
Research progress of lung organoids in infectious respiratory diseases
Li, ., Yang, W., Liu, X., Yang, K., Zhou, J., & Yang, X. (2025) · European Journal of Pharmacology · doi:10.1016/j.ejphar.2025.178201 · Cited by 5 · see citing works
Kim, S. H., Choi, S., Kim, S.-Y., Baek, J.-W., Park, C., & Kim, E.-M. (2025) · Environmental Toxicology and Pharmacology · doi:10.1016/j.etap.2025.104852 · Cited by 3 · see citing works
Chen, Q., Qi, H., & Guo, J. (2025) · Infection · doi:10.1007/s15010-025-02587-9 · Cited by 3 · see citing works
Understanding and manipulating morphogenetic processes to generate in vitro models of airways
Raasch, K., Henrot, P., Hadchouel, A., Zysman, M., & Dupin, I. (2025) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00015.2025 · Cited by 3 · see citing works
Human airway organoids as a versatile model to study BSL-4 virus replication and pathogenesis
Wälzlein, J.-H., Reusch, S., Ospina-Garcia, J., Olmer, R., Schneider, M. A., Klotz, L. V., Klotz, C., & Kummer, S. (2026) · Scientific Reports · doi:10.1038/s41598-026-45813-6 · Cited by 1 · see citing works
Zhang, W., Wang, T. W., Yao, M., Su, L., Xie, L., & Pan, P. (2026) · Open Research (University of Surrey) · doi:10.1002/viw.20250207 · Cited by 1 · see citing works
García‐Giménez, J. L., Seco-Cervera, M., Cánovas‐Cervera, I., Agúndez, A. B., Dolz-Andrés, E., Mondaray-Muñoz, G., Calpe-Fortea, R., Nácher-Sendra, E., Osca-Verdegal, R., Aparicio-Collado, J. L., et al. (2026) · Materials Today Bio · doi:10.1016/j.mtbio.2026.103194 · Cited by 1 · see citing works
Gellner, T. B., Chen, B., Raini, S. R., Jackson, M. S., Kraak, A. K., Petta, S. C., Paschini, M., Schnapp, L. M., Kim, C. F., & Liu, M. Y. (2026) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00356.2025 · Cited by 1 · see citing works
Endothelial Continuum and Capillary Specialization in Pulmonary Vascular Development
Ceas, A., Jimenez, A., Liu, Y., Niz, M. D., & Ellis, L. V. (2026) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.126.322483 · Cited by 1 · see citing works
The mechanical microenvironment and lung stem cell fate
Salazar, E. S. N., & Nelson, C. M. (2026) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2026.1836024
Angiogenesis in Lung Regeneration and Aging
Kyi, P., Scheer, M., Mammoto, T., & Mammoto, A. (2026) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.126.324410
New approach methodologies based prediction for pulmonary toxicity
Lee, J., Han, D., & Yang, S. (2026) · Toxicological Research · doi:10.1007/s43188-026-00376-6
- Authors
- Ament, A.-L., Heiner, M., Hessler, M. C., Alexopoulos, I., Steeg, K., Gärtner, U., Vazquez-Armendariz, A. I., & Herold, S.
- Affiliations
- University of Bonn (DE)
- Justus-Liebig-Universität Gießen (DE)
- Department of Medicine V (Internal Medicine, Infectious Diseases and Infection Control), Universities of Giessen and Marburg Lung Center (UGMLC), Giessen, Germany
- Venue
- American Journal of Respiratory Cell and Molecular Biology
- Publisher
- Oxford University Press (OUP)
- Published
- 2025-02-01
- Volume
- 72
- Issue
- 2
- Pages
- 124-132
- ISSN
- 1044-1549 (ISSN-L)
- 1535-4989
- OpenAlex
- W4402208231
- Cited by
- 17
- References
- 43
- Citations by year
- 2026: 8 · 2025: 9
- FWCI
- 4.94
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- American Journal of Respiratory Cell and Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Deutsche Forschungsgemeinschaft (KFO309/284237345, SFB-TR84/114933180, SFB1021/197785619)
- Hessisches Ministerium für Wissenschaft und Kunst
- Deutsches Zentrum für Lungenforschung (82DZL005B1)
- Cardio-Pulmonary Institute
- Topics
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Neonatal Respiratory Health Research
- Health Sciences › Medicine › Surgery › Transplantation: Methods and Outcomes
- Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
- Keywords
- pulmonary endothelial cells, endothelialization, microinjection, lung injury, influenza A virus, inflammatory cytokines, ICAM-1, alveolar epithelium, vascular endothelial cells
- MeSH terms
- Animals, Cell Differentiation, Cells, Cultured, Humans, Lung, Mice, Inbred C57BL, Models, Biological, Organoids, Pulmonary Alveoli, Intercellular Adhesion Molecule-1, Endothelial Cells, Mice, Lung Injury
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
