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. 2017 Nov 30;7:16675. doi: 10.1038/s41598-017-16856-7

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© The Author(s) 2017

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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Hepatocyte functionalities of hiHeps. (A) MRC5 cells were transduced with LV-5TFs or LV-control for 12 hr, and cultured until day 28. The hiHeps showed hepatic morphology. The scale bars represent 200 μm. (B) MRC5 cells were transduced with LV-5TFs or LV-control for 12 hr, and cultured until day 28. These cells were subjected to immunostaining with anti-ALB (green) and anti-AAT (red) antibodies. Nuclei were counterstained with DAPI (blue). The scale bars represent 20 μm. (C) The percentage of ASGR1-positive cells in LV-control- or LV-5TFs-transduced cells was examined by FACS. (D) The temporal ALB secretion capacity was examined by ELISA in MRC5 cells transduced with LV-5TFs. (E) The CYP1A2 and CYP3A4 activities were examined in MRC5 cells transduced with LV-5TFs, human iPS-Hepa, PHH 48 hr, and PHH 4 hr. The CYP1A2 and CYP3A4 activity levels in PHH 48 hr were taken as 1.0. *p < 0.05; **p < 0.01 (vs hiHeps). All data are represented as means ± SD (n = 3). PHH 48 hr: PHH cultured for 48 hr after plating; PHH 4 hr: PHH cultured for 4 hr after plating.