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. 2019 Jan 24;9:500. doi: 10.1038/s41598-018-36735-z

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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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Differentiation of primary NHBE cells from passaged cells grown in M3 (PnC-Ex-PLUS) expansion medium. NHBE cells expanded in M3 expansion medium to passage 1, 2, 4, 6 and 8 were cultured at ALI for 4 weeks in ALI differentiation media. The resulting 3D cultures were stained with H&E. (A) H&E staining of passaged NHBE cells (4 weeks ALI; X600 objective). (B) Trans-epithelial electrical resistance (TEER) of 3D cultured NHBE cells, 4 weeks ALI. Significance compared to first passage. **p < 0.01, ****p < 0.0001.