Fig. 2
Inhibition of T cells proliferation in the presence of co-culture with MSCLC or MSCHC. A CD3+ T cells were isolated from human PBMCs and stained with Celltrace Violet (CTV) proliferation dye to track T cell proliferation. Co-culture of T cells with MSCLC or MSCHC from Donor 1 to 6 was performed with CTV-stained T cells and anti-CD3/CD28 antibodies (Abs). B Representative histogram plots of CD3+ T cell proliferation in co-culture with MSCs, assessed by CTV dilution. C The extent of T cell proliferation was determined with T cell proliferative index (PI) calculated from their respective histogram by the following formula: PI = Log[FInd/MFIall]/Log[2], with MFIall = median fluorescence intensity of all viable T cells and FInd = peak fluorescence intensity of the viable non-divided cells. White bar indicates co-culture of MSCs harvested at low confluency (MSCLC) with T cells; light grey bar indicates co-culture of MSCs harvested at high confluency (MSCHC) with T cells; black bar indicates positive control (PC) containing anti-CD3/CD28-activated human CD3+ T cells without MSCs; dark grey bar indicates negative control (NC) containing naïve CD3+ T cells. The concentration of D IFN-ɣ and E IL-17A secreted into the co-culture media were measured by ELISA. Data shown were expressed as mean ± SEM of triplicates of separate experiments, representative of 6 donors. One-way ANOVA (Sidak’s multiple comparison test) was performed between selected pairs in each donor. *p ≤ 0.05; ** p ≤ 0.01 *** p ≤ 0.001; ns represents non-significant