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. 2018 Sep 17;8:13877. doi: 10.1038/s41598-018-31880-x

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© The Author(s) 2018

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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Influenza A virus replication in 3D printed and 2D cultured A549 cells. (A) Printed cell-laden alginate/gelatin constructs with varying Matrigel concentrations were infected with 4 × 10⁵ PFU of the Pan/99(H3N2) virus. Supernatants were collected at indicated time points and titrated on MDCK cells. (B) 2D cultured A549 monolayers were infected with the Pan/99(H3N2) virus at an MOI of 1, and collected supernatants were titrated on MDCK cells. (C) Metabolic activity of A549 cells was determined by XTT assays 24 h after infection of the 3D printed alginate/gelatin/Matrigel constructs with 10⁶ or 10⁷ PFU of Pan/99(H3N2). Values were calculated as X-fold induction of the lysis control. (D) Cytotoxicity resulting from infection with either 10⁶ or 10⁷ PFU of Pan/99(H3N2) was analysed by the LDH assay 24 h after infection. Data are depicted as percentage of the LDH amounts of cells in 3D printed constructs relative to the lysis control. Results are shown as mean ± SEM of three independent experiments.