Cell Growth Medium

[所属分类:培养基配方] [发布时间:2020-4-13] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Cell Growth Medium

培养基配方
Composition per 2250.0mL:
Eagle’s minimal essential medium
with Hanks’ salts (MEMH)...........................................1.0L
L 15 medium, modified Leibovitz.....................................1.0L
Fetal calf serum.................................................200.0mL
NaHCO3 solution...................................................50.0mL
pH 7.5 at 25°C
MEMH:
Composition per liter:
NaCl...............................................................8.0g
Glucose............................................................1.0g
KCl................................................................0.4g
CaCl2 ·2H2O .....................................................0.14g
MgSO4 ·7H2O.......................................................0.1g
KH2PO4 ...........................................................0.06g
Na2HPO4 ..........................................................0.05g
L- Isoleucine....................................................0.026g
L- Leucine ......................................................0.026g
L- Lysine........................................................0.026g
L- Threonine.....................................................0.024g
L- Valine.......................................................0.0235g
L- Tyrosine .....................................................0.018g
L- Arginine.....................................................0.0174g
L- Phenylalanine................................................0.0165g
L- Cystine.......................................................0.012g
L- Histidine......................................................8.0mg
L- Methionine.....................................................7.5mg
Phenol Red........................................................5.0mg
L- Tryptophan.....................................................4.0mg
Inositol..........................................................1.8mg
Biotin ...........................................................1.0mg
Folic acid........................................................1.0mg
Calcium pantothenate .............................................1.0mg
Choline chloride..................................................1.0mg
Nicotinamide......................................................1.0mg
Pyridoxal·HCl....................................................1.0mg
Thiamine·HCl ....................................................1.0mg
Riboflavin .......................................................0.1mg
Preparation of MEMH: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
L 15 Medium, Modified Leibovitz:
Composition per liter:
NaCl...............................................................8.0g
DL- Threonine......................................................0.6g
Sodium pyruvate....................................................0.6g
DL- Alanine........................................................0.5g
L- Arginine, free base.............................................0.5g
KCl................................................................0.4g
L- Asparagine·H2O.................................................0.3g
L- Histidine, free base............................................0.3g
L- Glutamine.......................................................0.3g
L- Isoleucine......................................................0.3g
L- Phenylalanine...................................................0.3g
L- Tyrosine........................................................0.3g
DL- Methionine.....................................................0.2g
DL- Valine.........................................................0.2g
Glycine............................................................0.2g
L- Serine .........................................................0.2g
Na2HPO4 , anhydrous ...............................................0.2g
CaCl2 , anhydrous..................................................0.1g
L- Cysteine, free base.............................................0.1g
L- Leucine·HCl....................................................0.1g
Streptomycin.......................................................0.1g
MgCl2 , anhydrous................................................0.094g
D- Galactose .....................................................0.09g
KH2PO4 ...........................................................0.06g
Gentamicin........................................................0.05g
L- Tryptophan.....................................................0.02g
Phenol Red........................................................0.01g
i-Inositol........................................................2.0mg
Choline chloride..................................................1.0mg
D- Calcium pantothenate...........................................1.0mg
Folic acid........................................................1.0mg
Nicotinamide......................................................1.0mg
Pyridoxine·HCl...................................................1.0mg
Thiamine monophosphate·2H2O......................................1.0mg
Riboflavin-5-phosphate ...........................................0.1mg
Penicillin G...................................................100,000U
pH 7.5 ± 0.2 at 25°C
Preparation of L 15 Medium, Modified Leibovitz: Add com-
ponents to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly. Filter sterilize. Store at 5°C.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ..........................................................7.5g
Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Aseptically combine 1.0L of sterile Ea-
gle’s minimal essential medium with Hanks’ salts (MEMH) and 1.0L
of sterile L 15 medium, modified Leibovitz. Mix thoroughly. Immedi-
ately prior to use, aseptically add 200.0mL of fetal calf serum and
50.0mL of NaHCO3 solution. Mix thoroughly. Aseptically distribute
into sterile containers.
Use: For the cultivation of mammalian HeLa or Vero tissue culture
cells to test the cytopathic effects of Escherichia coli.