Figure 3.
GMSCs regulate the immunophenotype and function of human monocyte leukemic cells (THP-1) during differentiation. (A–D): THP-1 cells (2 × 105) were differentiated on stimulation with PMA (10 ng/ml) in the absence or presence of IL-4 (100 ng/ml) or coculture with GMSCs in the transwell for 96 hours. Cells were stained with isotype-matched control IgGs (A), FITC-CD11a, and PE-conjugated CD14 (B), CD86 (C), or DC-SIGN (D) antibodies (A) and then analyzed by flow cytometry. THP-1 cells without any treatment were used as controls. (E–F): After coculture with GMSCs for 72 hours, THP-1 cells were stimulated with 0.1 and 1 μg/ml LPS for 4 hours (TNF-α) or 24 hours (IL-10), and the secretion of TNF-α (E) and IL-10 (F) in the supernatants was determined by enzyme-linked immunosorbent assay. (G): After coculture with GMSCs for 72 hours, THP-1 cells were stimulated with 0.1 and 1 μg/ml LPS for 1 hour, and the expression of NFκB p50 and p65 were determined by western blot, where the graphs represent the relative densities to the band of β-actin as the internal control. The results were representative of four independent experiments (mean ± SEM). *, p < .05; **, p < .01. Abbreviations: FITC, fluorescein isothiocyanate; GMSC, mesenchymal stem cells from human gingival; IL, interleukin; LPS, lipopolysaccharide; NFκB, nuclear factor kappa B; PE, phycoerythrin; PMA, phorbol 12-myristate 13-acetate; THP-1, human acute monocytic leukemia cell line; TNF, tumor necrosis factor.