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. 2017 Jul 24;7:6294. doi: 10.1038/s41598-017-06546-9

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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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Derivation of hepatocyte-like cells (HLC) from E-iPSCs. (A) First row phase contrast images of the differentiation stages, from undifferentiated stage the episomal induced pluripotent stem cells (E-iPSCs) to definitive endoderm (DE), then hepatic endoderm (HE) and finally hepatocyte-like cells (HLCs). Second row immunofluorescence-based staining of stage specific proteins overlapped with DAPI (staining of nucleus). Scale bar: 100 μm Alexa Flour 594 (red). (B) Immunofluorescence-based staining of HLC specific proteins AFP, ALB, HNF4α and A1AT. Scale bar: 100 μm Alexa Flour 594 (red). (C) Expression patterns of liver specific marker genes during HLC differentiation compared to fetal liver and primary human hepatocytes (PHH) are shown by quantitative real-time PCR (qPCR). Three biological replicates in technical triplicates of each sample were analyzed. The data were normalized to E-iPSCs. The standard deviation is depicted by the error bars.