Hydrogel-in-hydrogel live bioprinting for guidance and control of organoids and organotypic cultures

Anna Urciuolo (), Giovanni Giuseppe Giobbe, Yixiao Dong, Federica Michielin, Luca Brandolino, Michael Magnussen, Onelia Gagliano, Giulia Selmin, Valentina Scattolini, Paolo Raffa, Paola Caccin, Soichi Shibuya, Dominic Scaglioni, Xuechun Wang, Ju Qu, Marko Nikolic, Marco Montagner, Gabriel L. Galea, Hans Clevers, Monica Giomo, Paolo De Coppi and Nicola Elvassore ()
Additional contact information
Anna Urciuolo: University of Padova
Giovanni Giuseppe Giobbe: University College London
Yixiao Dong: ShanghaiTech University
Federica Michielin: University College London
Luca Brandolino: University of Padova
Michael Magnussen: University College London
Onelia Gagliano: University of Padova
Giulia Selmin: University College London
Valentina Scattolini: Città della Speranza
Paolo Raffa: Città della Speranza
Paola Caccin: University of Padova
Soichi Shibuya: University College London
Dominic Scaglioni: University College London
Xuechun Wang: ShanghaiTech University
Ju Qu: ShanghaiTech University
Marko Nikolic: University College London
Marco Montagner: University of Padova
Gabriel L. Galea: University College London
Hans Clevers: KNAW and University Medical Center
Monica Giomo: University of Padova
Paolo De Coppi: University College London
Nicola Elvassore: University College London

Nature Communications, 2023, vol. 14, issue 1, 1-14

Abstract: Abstract Three-dimensional hydrogel-based organ-like cultures can be applied to study development, regeneration, and disease in vitro. However, the control of engineered hydrogel composition, mechanical properties and geometrical constraints tends to be restricted to the initial time of fabrication. Modulation of hydrogel characteristics over time and according to culture evolution is often not possible. Here, we overcome these limitations by developing a hydrogel-in-hydrogel live bioprinting approach that enables the dynamic fabrication of instructive hydrogel elements within pre-existing hydrogel-based organ-like cultures. This can be achieved by crosslinking photosensitive hydrogels via two-photon absorption at any time during culture. We show that instructive hydrogels guide neural axon directionality in growing organotypic spinal cords, and that hydrogel geometry and mechanical properties control differential cell migration in developing cancer organoids. Finally, we show that hydrogel constraints promote cell polarity in liver organoids, guide small intestinal organoid morphogenesis and control lung tip bifurcation according to the hydrogel composition and shape.

Date: 2023
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DOI: 10.1038/s41467-023-37953-4

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