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. 2020 Dec 18;10:22294. doi: 10.1038/s41598-020-79448-y

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© The Author(s) 2020

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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Gene expression and protein immunofluorescence analysis of 3D sarcospheres scaffold-based models. Relative quantification of OCT-4, NANOG and SOX-2, stemness marker genes, SAOS-2 (a) and MG63 (b). Relative quantification of HIF-1a, NOTCH-1 and IL-6, CSCs niche-related genes, SAOS-2 (c) and MG63 (d). The graphs show the fold change expression of the genes relative to the scaffold-free sarcospheres at day 10 of culture (mean ± standard error; *p value ≤ 0.05; **p value ≤ 0.01; ***p value ≤ 0.001; ****p value ≤ 0.0001;). The panel shows a representative image of OCT-4 immunolocalization in SAOS-2 sarcorpheres MgHA/Coll model (e) and in HA model (f) scale bar 50 µm; SOX-2 immunolocalization in MG63 sarcorpheres MgHA/Coll model (g) and in HA model (h) scale bar 25 µm. Blu: cell nuclei; green: OCT-4; red: SOX-2.