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. 2023 Jul 26;13:12130. doi: 10.1038/s41598-023-39425-7

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© The Author(s) 2023

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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Development of monolayer model using cyst-enriched organoid as a cell source. (a) Plots of the TEER values for each day in culture. Differentiation was initiated from day 2. TEER values were corrected by their corresponding background well and area of insert membrane. (b) Papp of lucifer yellow in monolayer cultured with expansion medium for 2 days and differentiation medium with and without 3.0 mM VPA for 3 days. Data represents mean ± s.d. (n = 3, biological replicates). (c) Expression of differentiation marker (Si) and transporter and metabolic enzyme (Abcb1a, Abcb1b, and Cyp3a9) in monolayer culture with expansion medium for 2 days and differentiation medium with and without 3.0 mM VPA for 3 days. Data represents mean ± s.d. (n = 3, biological replicates). (d) Scheme of evaluation for expansion and differentiation periods. (e) Papp of lucifer yellow in monolayer culture with expansion medium for 2 and 3 days and differentiation medium for 2, 3, 4, and 5 days. Data represents mean ± s.d. (n = 3, biological replicates). (f) Expression of differentiation marker (Si) and transporter and metabolic enzyme (Abcb1a, Abcb1b, and Cyp3a9) in monolayer culture with expansion medium for 2 and 3 days and differentiation medium for 2, 3, 4, and 5 days. Data represents mean ± s.d. (n = 3, biological replicates). **p < 0.01, ***p < 0.001, ****p < 0.0001, Student t test. N Noggin, R R-spondin 1, A A83-01, Y Y-27632, g Gastrin, VPA valproic acid, TEER transepithelial electrical resistance, Papp apparent permeability coefficient.