@article{be3e1d3885ae4eb59357a6e3afc50a75,
title = "Polymer film-based microwell array platform for long-term culture and research of human bronchial organoids",
abstract = "The culture of lung organoids relies on drops of basement membrane matrices. This comes with limitations, for example, concerning the microscopic monitoring and imaging of the organoids in the drops. Also, the culture technique is not easily compatible with micromanipulations of the organoids. In this study, we investigated the feasibility of the culture of human bronchial organoids in defined x-, y-and z-positions in a polymer film-based microwell array platform. The circular microwells have thin round/U-bottoms. For this, single cells are first precultured in drops of basement membrane extract (BME). After they form cell clusters or premature organoids, the preformed structures are then transferred into the microwells in a solution of 50% BME in medium. There, the structures can be cultured toward differentiated and mature organoids for several weeks. The organoids were characterized by bright-field microscopy for size growth and luminal fusion over time, by scanning electron microscopy for overall morphology, by transmission electron microscopy for the existence of microvilli and cilia, by video microscopy for beating cilia and swirling fluid, by live-cell imaging, by fluorescence microscopy for the expression of cell-specific markers and for proliferating and apoptotic cells, and by ATP measurement for extended cell viability. Finally, we demonstrated the eased micromanipulation of the organoids in the microwells by the example of their microinjection.",
keywords = "Bronchi, bronchioli, Organoids, Microwells, Microthermoforming, Organoid fusion, (Organoid) microinjection, STEM-CELLS, LUNG, MECHANISMS, MODEL, REGENERATION, PLASTICITY, DISEASE, FUSION",
author = "D. Baptista and Z.T. Birgani and H. Widowski and F. Passanha and V. Stylianidis and K. Knoops and E. Gubbins and C. Iriondo and K.P. Skarp and R.J. Rottier and T.G. Wolfs and {van Blitterswijk}, C. and V. LaPointe and P. Habibovic and N.L. Reynaert and S. Giselbrecht and R. Truckenmuller",
note = "Funding Information: The authors thank Pinak Samal and Philipp Maurer from the MERLN institute for the design, fabrication and/or provision of the micromolds. This research was funded by the European Union, Horizon 2020 European Research Council Advanced Grant (project {\textquoteleft}ORCHESTRATE – Building complex life through self-organization: from organ to organism{\textquoteright}; ID 694801) and the Dutch Lung Foundation (Longfonds; no. 6.1.16.088). D. B. Z. T. B. F. P. E. G. C. v. B. V. L. P. H. S. G. and R. T. acknowledge financial support by the Dutch province of Limburg (program {\textquoteleft}Limburg INvesteert in haar Kenniseconomie/LINK{\textquoteright}; nos. SAS-2014-00837 and SAS-2018-02477). Z. T. B. P. H. S. G. and R. T. acknowledge the European Union, Interreg Flanders-The Netherlands (project {\textquoteleft}Biomat on microfluidic chip{\textquoteright}, no. 0433). E. G. C. v. B. P. H. S. G. and R. T. acknowledge the Gravitation Program (project {\textquoteleft}Materials-driven regeneration: Regenerating tissue and organ function with intelligent, life-like materials{\textquoteright}; no. 024.003.013) of the Dutch Research Council (Nederlandse Organisatie voor Wetenschappelijk Onderzoek; NWO). Funding Information: The authors thank Pinak Samal and Philipp Maurer from the MERLN institute for the design, fabrication and/or provision of the micromolds. This research was funded by the European Union, Horizon 2020 European Research Council Advanced Grant (project {\textquoteleft}ORCHESTRATE – Building complex life through self-organization: from organ to organism{\textquoteright}; ID 694801 ) and the Dutch Lung Foundation (Longfonds; no. 6.1.16.088 ). D. B., Z. T. B., F. P., E. G., C. v. B., V. L., P. H., S. G. and R. T. acknowledge financial support by the Dutch province of Limburg (program {\textquoteleft} Limburg INvesteert in haar Kenniseconomie /LINK{\textquoteright}; nos. SAS-2014-00837 and SAS-2018-02477 ). Z. T. B., P. H., S. G. and R. T. acknowledge the European Union , Interreg Flanders-The Netherlands (project {\textquoteleft}Biomat on microfluidic chip{\textquoteright}, no. 0433 ). E. G., C. v. B., P. H., S. G. and R. T. acknowledge the Gravitation Program (project {\textquoteleft} Materials-driven regeneration: Regenerating tissue and organ function with intelligent, life-like materials {\textquoteright}; no. 024.003.013 ) of the Dutch Research Council (Nederlandse Organisatie voor Wetenschappelijk Onderzoek; NWO). Publisher Copyright: {\textcopyright} 2023 The Authors",
year = "2023",
month = apr,
day = "1",
doi = "10.1016/j.mtbio.2023.100603",
language = "English",
volume = "19",
journal = "Materials today. Bio",
issn = "2590-0064",
publisher = "Elsevier BV",
number = "1",
}