Effect of Centipede grass on 3T3-L1 preadipocyte differentiation. 3T3-L1 preadipocytes were induced to differentiate as described in Materials and Methods. At day 4 or 6, cell viability and cytotoxicity were assessed in MTT and LDH assays. (A) 3T3-L1 preadipocytes was incubated with DMII media for 4 or 6 days and treated with different concentration of CG every day. Con, 3T3-L1 preadipocytes; DMII, fully differentiated adipocytes (0.5 mM 3-isobutyl-1-methylxanthine, 100 μM indomethason, 0.25 μM dexamethasone and 167 nM insulin); 10 μg/ml, fully differentiated adipocytes (DMII + 10 μg/ml CG); 100 μg/ml, fully differentiated adipocytes (DMII + 100 μg/ml CG). Cell viability was determined by MTT assay. Results represent the mean ± SD of three independent experiments. (B) LDH assay was performed to determine cytotoxicity during differentiation. Results represent the mean ± SD of three independent experiments. (C) 3T3-L1 preadipocytes were differentiated into adipocytes in DMII medium contained with or without different concentration of CG for 6 days. On day 6, cells were stained with Oil-red O to visualize lipid accumulation. (D) CG reduced TG content during differentiation of 3T3-L1 cells. 3T3-L1 preadipocytes were differentiated in the absence or presence of CG for 4 or 6 days, and the lipid accumulation was measured by triglyceride assay. Data are mean ± SD values of at least three independent experiments. (*) p < 0.05, (**) P < 0.01 compared with the differentiated adipocytes (DMII).