Cells were cultured in steroid-free, charcoal-dextran treated FBS (CDFBS, charcoal-dextran treatment removes steroid components) to observe the steroidal effect on the cultured muscle cells.
The present study adopted the novel track of using serum from castrated adult bovine as an alternative to CDFBS to observe steroidal effects during in vitro cell culture.
In addition, this study highlights the possibility of the use of castrated adult serum as an alternative to CDFBS in studying the effect of steroids.
Charcoal-dextran stripping of fetal bovine serum (CDFBS) was performed as previously described (Soto et al.
The seeding medium was then replaced with sample extracts diluted with 5% CDFBS in phenol red-free DMEM (Irvine Scientific, Santa Ana, CA, USA).
Water extracts (100 [micro]L in ethanol) were prepared by adding 5% CDFBS to a final volume of 10 mL and tested at five different concentrations to ensure a cell number near the half-maximal level ([M.sub.50]) of the 17[beta]-[E.sub.2] dose-response curve.
We removed sex steroids from FBS by charcoal-dextran stripping (CDFBS) (12).
The 5% CDFBS medium for proliferation assay consisted of plain DME, 4 mM L-glutamine, 2.24 g/L sodium hydrogen carbonate, and 5% CDFBS.