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. 2017 Apr 13;7:874. doi: 10.1038/s41598-017-00923-0

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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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AdMSC immunosuppressive activity. AdMSC immunosuppressive activity over T cell proliferation induced by anti-CD3 monoclonal antibody after 5 days of culture. AdMSC were cocultured with different concentrations with CFSE-labeled PBMC stimulated with anti-CD3 antibody. AdMSC:PBMC ratios were 1:10, 1:50 and 1:100 (n = 11). After 5 days, PBMC were intracellularly labeled with anti-CD3 PE-CY5 antibody (Becton, Dickinson and Company, USA) and analyzed by FACS (FACScalibur, BD Company, USA), to determine AdMSC suppressive activity over the T cell proliferation, in a cell-cell contact condition. To calculate the percentage of proliferation inhibition, the following formula was used, considering all conditions with anti-CD3 stimulus: [(% CD3 + CFSE^low cells- % CD3+ CFSE^low cells with AdMSC)/% CD3+ CFSE^low cells] * 100. The background of CFSE-positive unstimulated cells was lower than 1%. Note assays displaying high (>50%) or low (<50%) suppressive activity. Percentage of inhibition of T cell proliferation was compared usind AdMSC:PBMC different ratios. One-Way ANOVA. *p < 0.05.