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

• Published in: Nature communications Vol. 14; no. 1; p. 3128
• Main Authors: Urciuolo, Anna, Giobbe, Giovanni Giuseppe, Dong, Yixiao, Michielin, Federica, Brandolino, Luca, Magnussen, Michael, Gagliano, Onelia, Selmin, Giulia, Scattolini, Valentina, Raffa, Paolo, Caccin, Paola, Shibuya, Soichi, Scaglioni, Dominic, Wang, Xuechun, Qu, Ju, Nikolic, Marko, Montagner, Marco, Galea, Gabriel L., Clevers, Hans, Giomo, Monica, De Coppi, Paolo, Elvassore, Nicola
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30.05.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 13/1; 13/100; 13/106; 14/69; 147/3; 631/61/2035; 631/61/54/2295; 631/80/128; Bioprinting; Cell culture; Cell migration; Cell Polarity; Composition; Crosslinking; Differential geometry; Fabrication; Hepatocytes; Humanities and Social Sciences; Hydrogels; Hydrogels - chemistry; Lung; Mechanical properties; Morphogenesis; multidisciplinary; Organoids; Photon absorption; Photosensitivity; Regeneration; Science; Science (multidisciplinary); Self-assembly; Small intestine; Three dimensional printing; Tissue Engineering - methods
• ISSN: 2041-1723
• DOI: 10.1038/s41467-023-37953-4

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. Organ-like 3D cultures are advanced model system for biology and medicine limited by their uncontrolled cell self-assembly. Here, the authors develop a hydrogel-in-hydrogel bioprinting approach to dynamically control the growth landscape of a broad range of living 3D cell cultures.