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. 2020 Nov 20;22(1):77–91. doi: 10.1007/s10561-020-09867-8

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© The Author(s) 2020

Open AccessThis 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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Fig. 4 — Alizarin red S staining (ARS) assays of hADSCs after osteoinduction to determine the optimal time-span for exosomes. A: ARS staining of hADSCs incubated with PM, OM and different time-span of osteogenic induction exosomes (Exo^0d, Exo^1−14d and Exo^15−28d) on day 21. Scale bar = 200 μm. A’ The higher magnification panels of black box in Fig. 4A. The black arrow indicated the calcified nodules. B Quantification of ARS of hADSCs treated with or without exosomes. PM: proliferation medium; OM: osteogenic medium. ***p < 0.001 compared with PM group, ^ΔΔΔp < 0.001 compared with OM group, ^###p < 0.001 represents significant differences between compared groups. (Color figure online)