Fig. 3

Pimecrolimus exerts a suppressive effect on the immunomodulatory functions of hUCB-MSCs in vitro. Cells were treated with 100 ng/ml pimecrolimus during culture. a The immunosuppressive properties of pimecrolimus-treated hUCB-MSCs were determined using the MLR assay by measurement of co-cultured hPBMCs’ proliferation using the BrdU assay. b After coculture with hUCB-MSCs during the sensitization period (24 h), LAD-2 cells were challenged with anti-IgE. The degranulation rate of LAD-2 cells was assessed by detecting β-hexosaminidase in the culture medium. c The proliferation rate of LAD-2 cells was assessed using the BrdU assay.  d Expression levels of TNF mRNA in LAD-2 cells were determined using Quantitative Real-time PCR. e The expression of CD4 and IL-4 was determined using a flow cytometry analysis. Th2: Th2 cells, Pimecrolimus: Th2 cells treated with pimecrolimus, MSC: Th2 cells cocultured with MSCs, MSC + Pime.: Th2 cells cocultured with MSCs and pimecrolimus. f–h Expression levels of the GATA3 (f), STAT6 (g) and IL-13 (h) mRNAs in Th2 cells were determined using Quantitative Real-time PCR. In vitro experiments were performed in triplicate using hUCB-MSCs isolated from each donor (N = 3). All experiments shown in this figure were analyzed using the Kruskal–Wallis test with Dunn’s post hoc test. n.s: not significant, *P < 0.05 and **P < 0.01. The results are presented as the means ± SD