“…BM cells were isolated from C57BL/6 J female mice by conventional methods 35 and suspended at 1.0 × 10 6 cells/mL in DC culture medium composed of complete RPMI 1640, 10% heat-inactivated FCS (Invitrogen, CA, USA), 2.5 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer (Invitrogen), 1 mM sodium pyruvate (Invitrogen), 50 μM 2-mercaptoethanol (Invitrogen), 1% non-essential amino acids (NEAA, Invitrogen), 100 U/mL of penicillin, and 100 μg/mL of streptomycin (Sigma, MO, USA). The cells were cultured in the presence of 100 ng/mL of Flt-3L (R&D systems, MN, USA) and test extracts, fractions, and compounds were maintained at 37 °C in a 5% CO 2 atmosphere.…”