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. 2023 May 30;14:3128. doi: 10.1038/s41467-023-37953-4

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Brightfield time lap images (0, 8, 20 h) of a hydrogel-embedded tumor spheroid (day 6 post printing) growing within a cage of HCC-PEG pillars fabricated 1 day post organoid culture. Scale bar, 100 μm. b Fluorescent images of the tumor spheroid in c, showing different stages of cellular migration through the pillars day 7 post printing. Scale bars, 100 μm. c Young’s modulus measured by atomic force microscopy of 4-arm (black) or 8-arm (gray) HCC-PEG hydrogels photo-crosslinked at increasing laser power (300 or 500 μW) and wavelength (800 nm). Hydrogels were sequentially fabricated within the same Matrigel drop. Data are shown as mean ± s.d. of three independent replicates; all measurements performed are reported; unequal variance Student’s t-test was used; P < 0.05 was considered statistically significant. d Brightfield time lapse images of hydrogel-embedded tumor spheroid 5 or 7 days after bioprinting of 4-arm (upper panels) or 8-arm (lower panels) HCC-PEG pillars fabricated 1 day of organoid culture. Scale bars, 100 μm. e Quantification of nuclei detected out of the pillars of the hydrogel-embedded tumor spheroid 7 days after bioprinting of 4-arm (black) or 8-arm (gray) HCC-PEG pillars fabricated 1 day of organoid culture. Representative images of 3D reconstructions with xyz coordinates and 50 μm scale bar are shown. f Four-arm HCC-PEG hydrogel pillars were fabricated 1 day or 7 days post organoid culture (days 0 and 6 post printing, respectively) around a hydrogel-embedded growing tumor organoid. The brightfield images show the growth of the caged tumor spheroid at different time points. Scale bar, 100 μm. g Representative fluorescent image of tumor spheroid as in e at 14 days from first bioprinting step (day 15 of organoid culture). Scale bar, 100 μm. h Representative fluorescent images of cancer cells protruding through the bars of the first bioprinted hydrogel cage, invading the surrounding space and migrating through the pillars of the second fabrication step. Scale bars, 100 μm.