Comparison of Immunological Characteristics of Mesenchymal Stem Cells from the Periodontal Ligament, Umbilical Cord, and Adipose Tissue

Figure 5

Expression of HLA-G following IFN-γ treatment. (a) RNAs were extracted from PDLSCs, UC-MSCs, and AD-MSCs prior to and following activation with 10 ng/mL IFN-γ for 24 hours, and RT-PCR analysis of the HLA-G subtypes was performed. The primers used are listed in Table 1. Ribosomal protein S18 (RPS18) was used as a reference gene. This experiment was performed three times, and a representative figure is shown. (b) Culture supernatants of each MSC, treated or nontreated with 10 ng/mL for 48 hours, were harvested and subjected for secretory HLA-Gs by ELISA. Samples were from three independent experiments and were duplicated. No statistical difference was observed among groups. (c) Inhibitory role of HLA-G on activated T-cell proliferation. A T-cell proliferation assay was performed using CFSE-loaded PBMCs. Cells were activated with anti-CD3 and anti-CD28 beads and cocultured with UC-MSCs, AD-MSCs, or PDLSCs at an MSC : PBMC ratio of 1 : 10 in the presence of PBS, neutralizing anti-HLA-G antibody (anti-HLA-G Ab), or isotype control antibody (IgG2a). All experiments were performed independently at least three times. The figure shows a graph depicting the means and SEMs for the experiments. Flow cytometric profiles are presented in Supplementary Figure 3.

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