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. 2019 Jun 21;9:9027. doi: 10.1038/s41598-019-45195-y

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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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Human iLEC form lung epithelia in vivo. Human iLEC were co-transplanted subcutaneously into NOD-SCID animals with human embryonic stem cells. (a) Hematoxylin and eosin staining show teratomas containing all three germ layers, ectoderm, mesoderm and endoderm lineages. (b) Immunofluorescence characterization of the teratomas demonstrates presence of human iLEC-derived GFP+ lung epithelial cells expressing NKX2-1 and KRT8/18. Ciliated (FOXJ1+), Basal (P63+), Goblet (MUC5ac+) and Type II alveolar cell (proSPC+) were found lining the iLEC-derived epithelium. Apical expression of CFTR was also detected. White hatched represent GFP+ iLEC areas co-expressing specified markers.White arrows point to positive cells. Yellow arrows point to iLEC-derived (GFP+) mesenchyme (Vimentin+). (c) High magnification of hepatic/pancreatic ductal tissue (KRT19 + Hpd1+, white hatched area) and neuronal (Nestin+) cell clusters were human ES cell-derived. Donor (GFP+) stromal cells (Vimentin+) surround human ES-derived epithelial (panKRT + GFP−). Pink hatched represent GFP− hES-derived areas co-expressing specified markers.Teratoma experiments were replicated n = 2 times with four human iLEC lines. L; lumen. Scale bars represent 22–29 μm.