Figure 5

ADO production by mono- and co-cultures of myeloma cells with different components (osteoblasts, osteoclasts and stromal cells) of the BM niche. (A) Scheme of the co-culture system involving direct cell-cell contacts. (B) ADO generation (µmol/L) assayed in supernatants of mono- (□) and co-cultures (■) after adding NAD⁺ (CD38/CD203a/CD73-dependent), ATP (CD203a/CD73-dependent) or AMP (CD73-dependent) for 30 min at 37°C. Supernatants were collected and analyzed by HPLC. Variability (fold-increase) of ADO levels were detected after exposing osteoclasts (differentiated PBMC), osteoblasts (SAOS-2 cell line) and stromal cells (HS-5 cell line) to the LP-1 myeloma cells after adding NAD⁺, ATP or AMP. Values are the mean ± SD of triplicate determinations. All data are representative of at least three experiments.