DMEM培养基

DMEM培养基 DMEM composition, including our modification for suspension growth Componentconcentration (mg/L)Sigma #CaCl * 2HO50 in suspension; 250 on platesC790222 Fe(NO) * 9 HO0.1F8508332 MgSO4 (anhydrous)97.67M2643 KCl400P5405 NaCl6400S5886 NaHPO109S501124 L-Arginine * HCl84A3909 L-Cystine * HCl63C7777 L-Glutamine350; original 584G5763L-Glutamic acid88; original 0 or 7.35G5638Glycine30G6388 L-Histidine * HCl * HO42H95112 L-Isoleucine105I7383 L-Leucine105L1512 L-Lysine * HCl146L1262 L-Methionine30M2893 L-Phenylalanine66P5030 L-Serine42S5511 L-Threonine95T1645 L-Tryptophan16T0271 L-Tyrosine * 2Na97T1145 L-Valine94V6504 D-Ca pantothenate4P5155 Choline Chloride4C7527 Folic acid4F8758 i-Inositol7.2I7508 Niacinamide4N0636 Pyridoxine * HCl4P6280 Riboflavin0.4R9504 Thiamine * HCl4T1270 D-glucose4500; add more during suspension growthG7021 Phenol red * Na15P5530 Pyridoxal * HClP6155 110 mg/L sodium pyruvate can be added PBS Buffer for Washing Cells and Autoclaving the Bioreactor Amounts for 10 x solution 137mM NaCl(M = 58.44 g)80.06g/l 2.7mM KCl(M = 74.55 g) 2.01g/l 1.5mM KHPO(M = 136.09 g) 2.04g/l24 8mM NaHPO(M = 141.96 g)11.36g/l24 pH = 7.2 Additives for Suspension Growth in Bioreactor0.1 % (v/w)Pluronic acid F-68 265mg/lCaCl x 2 HO(1.8 mM; M = 147.02 g)22 50mg/lCaCl x 2 HO(170 µM; M = 147.02 g)for suspension22 +50mg/lHeparin (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.