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. 2023 Dec 4;17(6):064103. doi: 10.1063/5.0170606

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© 2023 Author(s).

All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

Published open access through an agreement with Politecnico di Milano18981

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FIG. 4. — (a) Design of the ECM-mediated-contact device. (i) Microfluidic channel layer is composed of one central channel, two adjacent culture channels, and two media channels. Every channel is separated from the adjacent one by two arrays of trapezoidal microposts. The layer comprises one self-sealing micro-guide to facilitate the manual insertion of a micro-needle. (ii) The tank layer is composed of a central channel to be aligned to the central channel of the microfluidic channel layer. (iii) Cross section with dimensions of the ECM-mediated-contact platform. (iv) Fabricated ECM device. Scale bar is 1 cm. (b) Seeding procedure of the ECM-mediated-contact device: (i) PHH-laden collagen–fibrin gel is injected through the inner lateral channels; after polymerization, (ii) collagen–fibrin gel is injected in the central channel containing the needle; (iii) the needle is removed, and the fibronectin coating (in light blue) is performed; then, after rinsing with the medium, (iv) endothelial cells are seeded within the lumen to generate a cylindrical vascular channel. (v) Cross-sectional area of the ECM-mediated-contact device, once all the passages to obtain the model have been performed. (c) Liver sinusoidal model generated with PHH and GFP-HUVEC within the (i) direct-contact device and the (ii) ECM-mediated-contact device. Pictures on day 3 (24 h after GFP-HUVEC seeding) and day 9 (end of the experiment) of the culture under static and dynamic conditions. Scale bars are 140 μm. (d) LDH release starts to decrease from day 5 in all conditions, indicating high cellular viability. Statistical analysis was performed using two-way ANOVA with the Tukey correction: ** p = 0.002, *** p < 0.001. ECM-S, ECM-mediated-contact device in static condition; ECM-D, ECM-mediated-contact device in dynamic condition. (e) High cellular viability is confirmed by albumin production and CYP3A4 activity levels. (i) Albumin production is higher in the ECM-mediated-contact device, as compared to the direct-contact device. The highest albumin production occurs on day 7 of the culture, and the production is higher in the ECM-mediated-contact device (static 24.29 ± 3.65 and dynamic 22.82 ± 3.08 μg/day/million cells) compared with the direct-contact device (static 11.31 ± 2.63 and dynamic 14.68 ± 3 μg/day/million cells). Statistical analysis was performed using two-way ANOVA with the Tukey correction: * p = 0.033 ** p = 0.002, *** p < 0.001. (ii) CYP3A4 levels are high in all conditions in both devices.