Fig. 4.

Effects of MMCs on development of MDSCs under hyperglycemic conditions. BM cells from femur and tibia of B6 mice mixed with or without MMCs in presence of mouse recombinant GM-CSF (10 ng/ml) at ratio of 80:1 under normal (N) (5 mM, 90 mg/dl) or high (H) (25 mM, 450 mg/dl) glucose conditions. Cells isolated for examination 7 days later. a Isolated cells two-color stained with specific mAbs against CD11b and Gr-1 for flow analyses. Four groups divided based on presence or absence of MMCs and glucose levels. Production of MDSCs from different cultured systems measured. Double-positive CD11b and Gr-1 cells represent MDSCs. Bar graph displays yield of MDSCs for each group. Data expressed as mean CD11b⁺/Gr-1⁺ cells ± 1 SD (*P < .05). b Cell surface molecules stained with specific mAbs against CD40, CD80, CD86, B7H1, MHC class II, and F4/80. Numbers represent percentage of positive cells. c Expression of arginase 1 and iNOS mRNA from MDSCs determined using qPCR (*P < .05). Data representative of three separate experiments. iNOS inducible nitric oxide synthase, MDSC myeloid-derived suppressor cell, MMC mouse mesangial cell