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. 2017 Aug 31;7:10113. doi: 10.1038/s41598-017-10708-0

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© The Author(s) 2017

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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The spheroid formation on chitosan film is associated with an increase in Apoptosis. MSCs were seeded at 2.5 × 10⁴/cm² without (Monolayer) or with chitosan coating (Spheroid), followed by calculation of cell numbers (a), Live/Dead staining (48 hr; Scale bar = 50 μM) (b), TUNEL assay (Scale bar = 10 μM) (c), and Annexin V/PI assay (d) at indicated time points. (e) Whole-cell lysates at 48 hr were analyzed by western blotting with specific antibodies against c-caspase-9 and c-caspase 3. β-tubulin was used as a loading control. The results are expressed as the mean ± standard deviation of three independent experiments, which is representative of MSCs from two individuals. ***p < 0.005 compared with monolayer cells.