The Human Mesenchymal Stromal Cell-Derived Osteocyte Capacity to Modulate Dendritic Cell Functions Is Strictly Dependent on the Culture System

Figure 1

Modulation of DC-induced T cell proliferation by hDOC. CD3⁺ T cells were cultured for 5 days either alone (negative control) or with allogeneic immature DC (positive control, +DC) or with both DC and hDOC. To evaluate the effect of detachment on the immunomodulatory function of hDOC, hDOC were either assayed with CD3⁺ and DC after detachment (+DC+Det-DOC) or left in adhesion (+DC+Adh-DOC). The ratio of seeded cells in coculture was the following: CD3⁺ T cell : DC : DOC = 10 : 1 : 1. (a) CFSE staining of CD3⁺ T cell alone (black line), CD3⁺ T cells with DC (black filled), CD3⁺ T cells with DC in presence of either Det-DOC (grey line), or Adh-DOC (dashed grey line); (b) histograms show the percentage of proliferating CD3⁺ T cells under the different conditions. (c) Effects of hDOC-derived extracellular matrix on CD3⁺ T cells proliferation. CD3⁺ T cells and immature DC were cultured for 5 days on hDOC-derived extracellular matrix, obtained by chemical lysis of Adh-DOC. (d) The same experiment as in (a) and (b) but using undifferentiated hMSC instead of hDOC. (e) The same experiment as in (a) and (b), performed in a 0.4 μm pore polycarbonate transwell system. Upper chamber: CD3⁺ T cells. Lower chamber: DC and hDOC. Histograms represent the mean ± SEM of the percentage of proliferating CD3⁺ T cells of 7 independent experiments. and , Bonferroni corrected.

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