DMEM/F-12, no phenol red

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	Seed 1x10 3 cells/well

Protocol tips
Seed 1x10 3 cells/well

Publication protocol

1x103 cells were counted and resuspended in 2 mL of mammosphere media (50 mL of Phenol-free DMEM (GIBCO, cat# 21041-025) supplemented with 1% L-glutamine, 1% penicillin/streptomycin, 1X B27 supplement minus Vitamin A (GIBCO, cat. #12587-010), 20 ng/ml rhEGF (R&D Systems, cat #236-EG-200) and 20 ng/ml bFGF (Invitrogen, Carlsbad, CA)) and cultured in a 6-well plate with an ultra-low attachment surface (Costar). The plate was left undisturbed for ten days in a 37°C incubator. Mammospheres were counted and measured using an inverted Axiovert 40 CFL microscope at 20X magnification.

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