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. 2019 Dec 30;9:20297. doi: 10.1038/s41598-019-55824-1

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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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Strategy of forming mixed organoids. (A) Schematic of the “mixed culture by direct sorting” procedure for forming one organoid derived from many stem cells. (B,C) Numbers of total (B) and Lgr5-enhanced green fluorescent protein (EGFP)^high (C) cells in organoids at Day 9 depend on the initial number of stem cells. Closed circles indicate individual organoids and black bars show their average (**P < 0.01, Tukey-Kramer). (D) Strategy of forming mixed organoids derived from two-color stem cells and their analysis. (E) Representative images of organoid growth. The scale bar shows 100 or 200 µm. The organoid was rotated because medium was added between days 4 and 5.