Fetus bovine serum fbs

CCs denuded from oocytes of ten to twenty patients were mixed together to reach an adequate amount for follow-up study and were transported immediately to the laboratory at room temperature. Cells were gently pipetted and centrifuged at 300 g for 5 min. The supernatant was discarded and the precipitate was resuspended in 500 μl hyaluronidase for 5 min. After addition of 2 ml Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12; Thermo Fisher Scientific Life Sciences, Massachusetts, USA) with 10% fetus bovine serum (FBS; Gibco, Grand Island, NE) and 1% penicillin-streptomycin (PS; Gibco, Grand Island, NE) solution, CCs were centrifuged at 300 g for 5 min and resuspended in 20 ml DMEM/F-12 medium with 10% FBS and 1% PS and plated for later use. As a positive control of ChemR23, THP-1 cell line (American Type Culture Collection, ATCC, Manassas, VA, USA) was thawed quickly in the water bath at 37°C and cultured in 1640 medium (Gibco, Grand Island, NE) with 10% FBS and 1% PS for further test. CCs and THP-1 were adherently cultured in T25 flask (Corning, NY, USA) at 37°C under 5% CO₂ in the air before use.

The SMMC 7721 human hepatoma and human cervical carcinoma HeLa cells were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in medium RPMI-1640 (Gibco-BRL, Carlsbad, CA, USA) supplemented with 10% fetus bovine serum (FBS) (Gibco-BRL) at 37°C with 5% CO₂. When the cells were 80% confluent, they were harvested by 0.25% trypsin digestion (Gibco-BRL, USA). The cells were then treated with serial concentrations of LMWSVP and untreated cells were used as controls.