Polymer film-based microwell array platform for long-term culture and research of human bronchial organoids.

Autor:	Baptista D; Department of Instructive Biomaterials, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Tahmasebi Birgani Z; Department of Instructive Biomaterials, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Widowski H; Department of Pediatrics, Maastricht University Medical Center (MUMC)+, Universiteitssingel 50, 6229 ER, Maastricht, the Netherlands.; GROW - School for Oncology and Reproduction, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Passanha F; Department of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Stylianidis V; Department of Respiratory Medicine, Maastricht University, Universiteitssingel 50, 6229 ER Maastricht, the Netherlands., Knoops K; Microscopy CORE Lab, Maastricht MultiModal Molecular Imaging Institute - M4i, Maastricht University, Universiteitssingel 50, 6229 ER, Maastricht, the Netherlands., Gubbins E; Department of Instructive Biomaterials, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands.; Department of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Iriondo C; Department of Pediatric Surgery/Cell Biology, Erasmus (University) Medical Center Rotterdam - Sophia Children's Hospital, Doctor Molewaterplein 40, 3015 GD, Rotterdam, the Netherlands., Skarp KP; Department of Pediatric Surgery/Cell Biology, Erasmus (University) Medical Center Rotterdam - Sophia Children's Hospital, Doctor Molewaterplein 40, 3015 GD, Rotterdam, the Netherlands., Rottier RJ; Department of Pediatric Surgery/Cell Biology, Erasmus (University) Medical Center Rotterdam - Sophia Children's Hospital, Doctor Molewaterplein 40, 3015 GD, Rotterdam, the Netherlands., Wolfs TG; Department of Pediatrics, Maastricht University Medical Center (MUMC)+, Universiteitssingel 50, 6229 ER, Maastricht, the Netherlands.; GROW - School for Oncology and Reproduction, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., van Blitterswijk C; Department of Instructive Biomaterials, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands.; Department of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., LaPointe V; Department of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Habibović P; Department of Instructive Biomaterials, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Reynaert NL; Department of Respiratory Medicine, Maastricht University, Universiteitssingel 50, 6229 ER Maastricht, the Netherlands.; School of Nutrition and Translational Research in Metabolism (NUTRIM), Universiteitssingel 40, Maastricht University, Maastricht, the Netherlands., Giselbrecht S; Department of Instructive Biomaterials, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands., Truckenmüller R; Department of Instructive Biomaterials, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands.
Jazyk:	angličtina
Zdroj:	Materials today. Bio [Mater Today Bio] 2023 Mar 14; Vol. 19, pp. 100603. Date of Electronic Publication: 2023 Mar 14 (Print Publication: 2023).
DOI:	10.1016/j.mtbio.2023.100603
Abstrakt:	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. Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Roman Truckenmueller reports a relationship with 300MICRONS GmbH that includes: board membership and equity or stocks. Stefan Giselbrecht reports a relationship with 300MICRONS GmbH that includes: board membership and equity or stocks. (© 2023 The Authors.)
Databáze:	MEDLINE
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