At this point, medium was changed to defined N2B27 medium (Invitrogen) lacking bFGF but including either 100 ng/ml BMP2 (PeproTech) or BMP4 (R&D Systems, Minneapolis, MN, USA, www.rndsystems.com) for a period of five days or a combination of 10 ng/ml BMP4 and 10 µM SB431542 (a TGFβRI inhibitor, Sigma-Aldrich) for 7 days.
This medium included KH2PO4, 6; Na2.7O4, 2.7; and glucose, 34 g/l.
The culture medium included Dulbecco's modified Eagle's medium (DMEM, Gibco, USA) with 15% fetal bovine serum (Gibco, USA).
The examined phenotypic traits performed in BHI medium included absorption of bromothymol blue dye, bioluminescence, colony morphology, and growth rate.
One medium included the complete range of steroids, including ample amounts of androgen and growth factors.
For growth curve analyses, the galactose medium included 50 μg/ml uridine.
The control (mock) liquid medium included the same amount of the solvent dimethylsulfoxide (0.04%).
For 2 3 days post-plating the medium included 15%% heat inactivated fetal calf serum.
In contrast, N-glycoproteins detected in the MCF-7 medium included CPA4, AAT, HP, and HSC70).
Interestingly, the group of reduced genes in the minimal medium included genes involved in nucleic acid metabolism (mainly adenine synthesis), although the medium included adenine, and none of the adenine pathway genes is mutated in this strain.
