... analytes, containing 1, and 10 nM of icariin, icaritin and desmethylicaritin; 5, 30 and 60 nM of icariside I and 71 icariside II, respectively and internal standard in pooled rat sera, were placed ... using lipofectamine Sixteen hours posttransfection, the cells were washed, trypsinized and split at ratio of in 10, in 100 and in 1000 into selective cell culture media containing 100 μg/mL of hygromycin ... fetal bovine serum and 10 μg/mL insulin in a humidified atmosphere of 5% carbon dioxide in air at 37°C 53 2.3 ERα and ERβ cell- based bioassays 2.3.1 Establishment of ERα and ERβ stable cells (from...