Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates

Here we present a protocol to engineer apical-out airway organoids (AOAOs) directly from human airway basal stem cells (hABSCs) using suspension culture of hABSC aggregates on a cell-repellent surface. We describe steps to produce spherical AOAOs with homogenous presentation of exterior-facing motil...

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Autores principales: Wijesekara, Piyumi, Patel, Kian Z., Otto, Ellen L., Campbell, Phil G., Ren, Xi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10025264/
https://www.ncbi.nlm.nih.gov/pubmed/36917607
http://dx.doi.org/10.1016/j.xpro.2023.102154
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author Wijesekara, Piyumi
Patel, Kian Z.
Otto, Ellen L.
Campbell, Phil G.
Ren, Xi
author_facet Wijesekara, Piyumi
Patel, Kian Z.
Otto, Ellen L.
Campbell, Phil G.
Ren, Xi
author_sort Wijesekara, Piyumi
collection PubMed
description Here we present a protocol to engineer apical-out airway organoids (AOAOs) directly from human airway basal stem cells (hABSCs) using suspension culture of hABSC aggregates on a cell-repellent surface. We describe steps to produce spherical AOAOs with homogenous presentation of exterior-facing motile cilia and of tunable sizes. We then detail procedures to analyze AOAO cellular composition via wholemount staining and assess cilia motility via 3D AOAO rotation upon Matrigel embedding. The protocol offers an effective model for investigating human airway pathophysiology. For complete details on the use and execution of this protocol, please refer to Wijesekara et al. (2022).(1)
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spelling pubmed-100252642023-03-21 Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates Wijesekara, Piyumi Patel, Kian Z. Otto, Ellen L. Campbell, Phil G. Ren, Xi STAR Protoc Protocol Here we present a protocol to engineer apical-out airway organoids (AOAOs) directly from human airway basal stem cells (hABSCs) using suspension culture of hABSC aggregates on a cell-repellent surface. We describe steps to produce spherical AOAOs with homogenous presentation of exterior-facing motile cilia and of tunable sizes. We then detail procedures to analyze AOAO cellular composition via wholemount staining and assess cilia motility via 3D AOAO rotation upon Matrigel embedding. The protocol offers an effective model for investigating human airway pathophysiology. For complete details on the use and execution of this protocol, please refer to Wijesekara et al. (2022).(1) Elsevier 2023-03-13 /pmc/articles/PMC10025264/ /pubmed/36917607 http://dx.doi.org/10.1016/j.xpro.2023.102154 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Wijesekara, Piyumi
Patel, Kian Z.
Otto, Ellen L.
Campbell, Phil G.
Ren, Xi
Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates
title Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates
title_full Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates
title_fullStr Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates
title_full_unstemmed Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates
title_short Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates
title_sort protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10025264/
https://www.ncbi.nlm.nih.gov/pubmed/36917607
http://dx.doi.org/10.1016/j.xpro.2023.102154
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