Figure 4.

Migratory Ability and Immunomodulatory Capability Assays with Engineered MSCs

(A) The cell migration of MSCs given different treatments (infected with empty-LV, IL-25-LV, CX3CR1-LV, or CX3CR1&IL-25-LV) was examined by Transwell migration assays after the cells were plated in the upper chamber, CX3CL1 was added in the bottom chamber, and the system was incubated for 24 h. Scale bars, 20 μm. (B) Cells were counted in 5 randomly selected fields, and cell counts were averaged. (C) The production of IL-12/IL-23 in LPS-activated APCs was detected after a 12-h exposure to the supernatants of MSCs infected with different types of LVs or the aforementioned supernatant supplemented with an anti-IL-25 mAb or rat IgG (2 μg/mL). (D) Naive CD4⁺ T cells magnetically purified from spleens harvested from C57BL/6 mice were cultured with the aforementioned treated medium from APCs for 96 h. (E) Representative flow cytometry plots are shown after gating on live CD4⁺ T cells, followed by gating on CD4⁺ IFN-γ⁺ cells or CD4⁺ IL-17A⁺ cells. ST, supernatant. For cell experiments, five samples were analyzed per condition, and the experiments were performed in triplicate. Values are expressed as the mean ± SEM *p < 0.05; **p < 0.01; ***p < 0.001.