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. 2021 Feb 5;12:111. doi: 10.1186/s13287-020-02091-x

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© The Author(s) 2021

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — ASC-EVs protect neurons from ischemic injury. a Representative microscopic image of morphology of ASCs after osteogenesis, adipogenesis, and chondrogenesis. ALP for osteogenesis, Oil Red O staining for adipogenesis, and Alcian Blue staining for chondrogenesis (× 400). b Immunofluorescence of MAP2 expression in ASCs (× 400). c CCK-8 assay detection of cell viability of OGD/RP-induced neurons incubated in ASC-CM and EV-free CM, or controls. d CCK-8 assay detection of OGD/RP-induced neurons upon co-culture with ASCs and GW4869-treated ASCs, or controls. e Representative of EVs from ASCs under TEM (bar = 100 nm). f Quantification of size distribution of EVs under NTA. g Western blot analysis of specific EV-surface protein expression. h Representative images of neurons internalizing Dil-label EVs (red) and DAPI-labeled neuron nucleus (blue) (× 400). i CCK-8 assay detection of neuron viability upon treatment of ASC-EVs. j Flow cytometry detection of apoptosis upon ASC-EV treatment. k Western blot analysis of caspase 3, cleared caspase 3, Bax, and Bcl-2. C/T refers to the cleaved caspase 3/total caspase 3. l Representative macroscopic images of cerebral infraction volume with TCC staining. m NeuN immunofluorescence of hippocampus upon treatment of ASC-EVs, PBS, and sham operation (× 400). n Representative images of TUNEL staining of hippocampus apoptosis upon treatment of ASC-EVs, PBS, and sham operation (× 200). *p < 0.05, **p < 0.01, and ***p < 0.001. Measurement data were presented as mean ± standard deviation. The data among multiple groups were analyzed by ANOVA