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. 2020 Dec 14;10:21900. doi: 10.1038/s41598-020-79082-8

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Forskolin-induced ion channel activity is more prominent in ECO then ICO. A representative Ussing experiment (A) shows CFTR stimulation by forskolin (FSK, 5 μM) in ECO (blue line) and ICO (red line) organoids. GlyH-101 inhibits the CFTR transporter channel specifically. Ca²⁺-activated Cl-channels (CaCC) are stimulated by UTP and inhibited by CaCC inhibitor T16inh-A01 (arrows) demonstrating the presence of functional CFTR and Ca²⁺-activated Cl-channels in these organoids. Functionality of CFTR in 3D grown organoids was shown by FIS. Representative pictures of five individual calcein green-labeled ECO (B) and ICO (C) before (t = 0) and after (t = 120) forskolin stimulation show the swelling in time. Qualification of organoid size in time showed that ECO swell significantly larger upon forskolin-activation than ICO (D) (*p < 0.0001).