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. 2019 Nov 5;9:16006. doi: 10.1038/s41598-019-52516-8

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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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Long-term storage and suspension culture of CMCs. (a) The morphology of cryopreserved CMCs post-thaw and after three passages in ABC medium. (b) The survival rate of cryopreserved CMCs post-thaw. (c) The percentages of MESP1⁺ and Ki67⁺ cells in cryopreserved CMCs after three passages in ABC medium as assessed by flow cytometry. (d) The morphology of generated CMC-spheroids in ABC medium at day 4 of suspension culture. (e) The percentage of MESP1⁺ cells after four passages of CMCs in suspension culture as evaluated by flow cytometry. (f,g) The calculated total cell count and expansion fold after four passages of CMCs in suspension culture condition. Scale bars: 200 µm. All data: mean ± SD.