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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 at ALI with three different culture conditions. NHBE cells at passage two were cultured at ALI for 4 weeks in different growth media. The resulting 3D cultures were stained with Hematoxylin and Eosin (H&E). (A) H&E staining of NHBE layers grown from cells cultured in M1 + ROCK (BEGM + ROCK), M2 (PnC Ex) or M3 (PnC-Ex-PLUS). (B) TEER measurements of epithelial layers grown in ALI from cells expanded in M1 + ROCK, M2 or M3 media. ****p < 0.0001.