Opi media supplement

Mono Mac 6 cells (DSMZ ACC 124)^{25 (link)}, a human monocyte-derived cell line exhibiting a mature monocyte phenotype, were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 mg/ml streptomycin, 1% non-essential amino acids (Life Technologies, Carlsbad, CA, USA), and 1% OPI media supplement (Sigma-Aldrich, St. Louis, MO, USA). THP-1 cells (ATCC TIB-202)^{29 (link)}, another human monocyte-derived cell line, were grown and maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 mg/ml streptomycin. These cells were grown at 37 °C in 5% CO2 in a humidified incubator.

The RAW264.7 (ATCC TIB-71) [18 (link)] and THP-1 cells (ATCC TIB-202) [20 (link)] were grown and maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 mg/ml streptomycin; and Mono Mac 6 cells (DSMZ ACC 124) [19 (link)] were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 mg/ml streptomycin, 1% non-essential amino acids (Life Technologies), and 1% OPI media supplement (Sigma-Aldrich). These cells were grown at 37°C in 5% CO2 in a humidified incubator. To differentiate THP-1 cells into macrophages, THP-1 cells were treated with phorbol-12-myristate-13-acetate (PMA) at the concentration of 100 nM for 72 hours. The cells were incubated in growth medium without PMA for an additional 24 hours, and used in the study. Normal human monocytes were purchased from commercially available products (Promocell, Heidelberg, Germany). The cells were differentiated into macrophages using a commercially available medium (M1-Macrophage Generation Medium DXF; Promocell) in accordance with the manufacturer's protocol, and used in the study. The procedure for preparation of the purified AP-PG from A. pullulans cultured fluid was described elsewhere [15 (link)]. Curdlan derived from Alcaligenes faecalis was obtained from commercially available products (Sigma-Aldrich).