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. 2017 Oct 11;7:12984. doi: 10.1038/s41598-017-13326-y

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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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Characterization of rat mesenchymal stem cells (rMSCs) from bone marrow with DIC microscopic picture; rMSCs stained with CD44 and CD90 antibody conjugated with TRITC and FITC; tri-lineage differentiation capability into chondrogenic lineage after 3D aggregation culture for 21 day, osteogenic lineage after 21 days, and adipogenic lineage after 21 days (scale bar: 50 μm) (A). The rMSC aggregate diameter based on cell number, with approximately 2,000 rMSCs used to form aggregates with diameters between 250 and 300 μm (B). Characterization of osteogenic differentiation of rMSC aggregates over 21 day, with 3D aggregates analyzed by immunostaining with collagen type I antibodies for collagen synthesis and by Alizarin Red S staining for mineralization (C). Image J software was used to quantify the positively stained area (scale bar: 50 μm).