Fig. 3

Effect of monocyte-ASC coculture on levels of monocyte subsets, COX-2, PGE2, and EP4. First, monocytes from healthy donors and sepsis patients were cultured alone or cocultured directly with ASCs. The percentage of CD14⁺⁺CD16⁺ (a), CD14⁺CD16⁺⁺ (b), and CD14⁺⁺CD16^– (c) monocytes in the total monocyte population were determined via flow cytometry. Box and whisker plots represent median (lines within boxes), interquartile range (bounds of boxes), and error bars (upper and lower range); n = 25 for healthy donors and n = 23 for sepsis patients. ***p < 0.001. Then, ASCs and monocytes from sepsis patients were cultured alone, cocultured directly, or cocultured via Transwell for 24 h. Culture supernatants from the above wells were harvested for quantification of PGE2 via ELISA (d). Lysates from different groups were analyzed for COX-2 and EP4 levels via Western blotting. Representative blots and normalized COX-2 levels (e) and EP4 (f) are shown. β-actin was used as a protein-loading control. Data are expressed as mean ± SEM; n = 5 per group. *p < 0.05, **p < 0.01. ASC adipose-derived mesenchymal stem (stromal) cell, COX-2 cyclooxygenase-2, EP4 prostaglandin E2 receptor 4, MO monocytes, PGE2 prostaglandin E2