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. 2019 Apr 15;9:6087. doi: 10.1038/s41598-019-42493-3

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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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Culture and characterization of primary human CEnCs. (A) Schematic diagram depicting the dual media culture system used in this study, from the procurement stage through to the different phases of processing including isolation, stabilization, expansion and passaging. (B) Representative image of a confluent culture of primary human CEnCs at the third passage showing the homogeneous cellular morphology. Cultures of human CEnCs were characterized for expression of (C) Na⁺/K⁺-ATPase, and (D) ZO-1 by immunocytochemistry, as well as their expression of (E) Tag 1A3 - CD166, and Tag 2A12 - PRDX-6 and by flow cytometric analysis of live CEnCs. Scale bar: B 100 µm; C and D 50 µm.