1、DMEM composition,including our modification for suspension growthComponent concentration (mg/L) Sigma #CaCl2 * 2H2O 50 in suspension; 250 on plates C7902Fe(NO3)3 * 9 H2O 0.1 F8508MgSO4 (anhydrous) 97.67 M2643KCl 400 P5405NaCl 6400 S5886NaH2PO4 109 S5011L-Arginine * HCl 84 A3909L-Cystine * HCl 63 C77
2、77L-Glutamine 350; original 584 G5763L-Glutamic acid 88; original 0 or 7.35 G5638Glycine 30 G6388L-Histidine * HCl * H2O 42 H9511L-Isoleucine 105 I7383L-Leucine 105 L1512L-Lysine * HCl 146 L1262L-Methionine 30 M2893L-Phenylalanine 66 P5030L-Serine 42 S5511L-Threonine 95 T1645L-Tryptophan 16 T0271L-T
3、yrosine * 2Na 97 T1145L-Valine 94 V6504D-Ca pantothenate 4 P5155Choline Chloride 4 C7527Folic acid 4 F8758i-Inositol 7.2 I7508Niacinamide 4 N0636Pyridoxine * HCl 4 P6280Riboflavin 0.4 R9504Thiamine * HCl 4 T1270D-glucose 4500; add more during suspension growth G7021Phenol red * Na 15 P5530Pyridoxal
4、* HCl P6155110 mg/L sodium pyruvate can be addedPBS Buffer for Washing Cells and Autoclaving the Bioreactor Amounts for 10 x solution137 mM NaCl (M = 58.44 g) 80.06 g/l2.7 mM KCl (M = 74.55 g) 2.01 g/l1.5 mM KH2PO4 (M = 136.09 g) 2.04 g/l8 mM Na2HPO4 (M = 141.96 g) 11.36 g/lpH = 7.2Additives for Suspension Growth in Bioreactor0.1 % (v/w) Pluronic acid F-68265 mg/l CaCl2 x 2 H2O (1.8 mM; M = 147.02 g)50 mg/l CaCl2 x 2 H2O (170 M; M = 147.02 g) for suspension50 mg/l Heparin (Na+-salt) OR 300 mg/ L dextran sulfateDialyze 500 ml FBS 3-times against 10 l PBS at 4 C with a tubing cut off of 1 kDa.
