Abstract
Organoids are a powerful model system to explore the role of mechanical forces in sculpting emergent tissue cytoarchitecture. The modulation of the mechanical microenvironment is most readily performed using synthetic extracellular matrices (ECM); however, such materials provide passive, rather than active force modulation. Actuation technologies enable the active tuning of mechanical forces in both time and magnitude. Using such instruments, our group has shown that extrinsically imposed stretching on human neural tube organoids (hNTOs) enhanced patterning of the floor plate domain. Here, we provide a detailed protocol on the implementation of mechanical actuation of organoids embedded in synthetic 3D microenvironments, with additional details on methods to characterize organoid fate and behavior. Our protocol is easy to reproduce and is expected to be broadly applicable to investigate the role of active mechanics with in vitro model systems.
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Acknowledgments
This work was supported by the FWO grants G087018N, G0ACA24N and I009718N, FWO postdoctoral fellowship 1217220 N, FWO doctoral scholarships 11K722N and 11M5323N, Interreg Biomat-on-Chip grant and Vlaams-Brabant and Flemish Government co-financing, KU Leuven grants C14/17/111, C32/17/027, IDN/22/012 and IDN/20/007, King Baudouin Foundation grant J1810950-207421 and an Allen Distinguished Investigator Award, a Paul G. Allen Frontiers Group advised grant of the Paul G. Allen Family Foundation.
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Sgualdino, F., Mattolini, L., Jimenez, B.D., Patrick, K., Abdel Fattah, A.R., Ranga, A. (2024). Mechanical Actuation of Organoids in Synthetic Microenvironments. In: Sumbalova Koledova, Z. (eds) 3D Cell Culture. Methods in Molecular Biology, vol 2764. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3674-9_15
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DOI: https://doi.org/10.1007/978-1-0716-3674-9_15
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