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. 2019 Sep 19;9:13557. doi: 10.1038/s41598-019-49412-6

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© The Author(s) 2019

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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Gene expression analysis in hepatocyte-like cells (HES2 line). In all panels, the levels of the target mRNA are normalized to the expression level of the 28S rRNA. (A) Real-time PCR of genes involved in metabolic and cell cycle processes in hepatocyte-like cells obtained following hESCs differentiation at uniform 21% O₂ concentration (control chip). No statistical difference is observed between the right side and the left side. Data, normalized to the average of the right side, are shown as mean ± SEM. n = 3. Real-time PCR of genes involved in metabolic and cell cycle processes in hepatocyte-like cells grown in a stable O₂ gradient generated by the microfluidic device. (B) GLUL (glutamine synthetase), CTNNB (beta-catenin) and CCND1 (cyclin D1) show significantly higher expression in low pO₂. (C) CPS-1 (carbamoyl-phosphate synthetase 1), ALB (albumin), MKI67, CCNA (cyclin A) show significantly higher expression in high pO₂. Data, normalized to the average of 21% O₂, are shown as mean ± SEM. n = 9. p* < 0.001 and p§ < 0.0001 respect to 21% O₂.