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. 2017 Aug 25;7:9408. doi: 10.1038/s41598-017-09686-0

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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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GBM microtumours embedded in Matrigel, formed by primary cells isolated from a clinical sample. (a) Static culture of GBM microtumours. (b) Perfusion culture of GBM microtumours. GBM microtumours were formed by embedding primary cells in Matrigel, and then the 3D structures were extracted as described in Materials and Methods, followed by staining with astrocyte differentiation marker glial fibrillary acidic protein (GFAP, red fluorescence) and pluripotency marker Nanog (green fluorescence). Scale bar: 100 µm. Quantification of fluorescent cells shown in bar graph.