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. 2019 Nov 14;9:16819. doi: 10.1038/s41598-019-53418-5

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© The Author(s) 2019

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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Increasing number of SSEA-3-positive cells after OGD in vitro and the presence of SSEA-3-positive cells after OGD-PBMCs administration in vivo. (A) Representative figures of octamer-binding transcription factor 3/4 (Oct3/4) (red)/stage-specific embryonic antigen-3 (SSEA-3) (green)/DAPI (blue) triple labelling of NTERA-2 cells, as examined by confocal microscopy. Scale bars, 20 μm. (B, C) Expression of SSEA-3 using both the human (B) and rat (C) OGD-PBMCs by flow cytometric analyses. The x-axis represents forward scatter and y-axis represents the SSEA-3-FITC channel. SSEA-3-positive cells in the OGD-PBMCs were more prominent than those in the normoxic PBMCs by unpaired t-test. **P < 0.01. (each N = 6). (D) The two point time series of expression of SSEA-3-positive cells in the human OGD-PBMCs by flow cytometric analyses by two-way ANOVA. (each N = 6). (E) SSEA-3-positive cell recruitment in the sham-operated and ischemic cerebral cortices at days 10 (i.e., administration after 3 days) and 28 (i.e., administration after 21 days) after ischemia. A bar graph represents the total number of SSEA-3-positive cells. Statistical analyses were performed by one-way ANOVA. A secondary-only antibody control confirms its specificity. Scale bars, 20 μm. *P < 0.05. (N = 28).