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The cells were then inoculated into N2 or B27-supplemented RPMI-1640/1% methylcellulose medium without serum at a density of 1x105 cells/well in ultra low attachment six-well plates N2 medium consists of 2X DMEM/F12 supplemented with progesterone (20 nM), putrescine(100 μM), sodium selenite (30 nM), transferrin (25 μg/ml),insulin (20 μg/ml), and human recombinant epidermal growth factor (EGF; 10 ng/ml) and basic fibroblast growth factor (bFGF; 10 ng/ml) mixed with an equal volume of 2% methylcellulose.
% \ B2 F3 u' T$ X EGF,bFGF是和DMEM/F12混合后浓度为10ng/ml,然后再和等量的2%methylcellulose混合?还是和等量的2%methylcellulose混合后最终浓度是10ng/ml? |
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