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US 7,923,245 B2Granted
Medium for ES culturing
OwnerMiho Furue, Tetsuji Okamoto +1inventors
InventorsMiho Furue, Tetsuji Okamoto, Makoto Asashima
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Abstract
The present invention discloses a medium for a serum-free medium capable of culturing ES cells for a long period while maintaining their undifferentiated state without using feeder cells, and a basal medium for producing the medium thus described. The basal medium of the present invention is characterized by that it has composition shown by Table I. Further, the present invention discloses a medium for ES cells produced with the basal medium.
Claims5
A medium for culturing embryonic stem (ES) cells comprising:
- (a) insulin, transferrin, 2-mercaptoethanol, 2-ethanolamine, sodium selenite, oleic acid which has formed a complex with fatty acid-free bovine serum albumin, and LIF (leukemia inhibitory factor;
- and (b) the basal medium comprising the composition shown by the following Table III.
- TABLE III
- Components
- Concentration (mg/L)
- L-alanine
- 1.78~2.67
- L-arginine
- 40~60
- L-arginine HCl
- 75.8~113.7
- L-asparagine H2O
- 13.002~19.503
- L-Asparatic acid
- 6.66~9.99
- L-cysteine HCl•H2O
- 7.024~10.536
- L-cystine 2HCl
- 38.058~57.087
- L-glutamic acid
- 6.94~10.41
- L-glutamine
- 439.72~659.58
- Glycine
- 15.5~23.25
- L-histidine
- 3~30
- L-hydroxyproline
- 4~6
- L-isoleucine
- 52.748~79.122
- L-leucine
- 54.58~81.87
- L-lysine HCl
- 73.74~110.61
- L-methionine
- 15.896~23.844
- L-phenylalanine
- 30.392~45.588
- L-proline
- 10.9~16.35
- L-serine
- 24.9~37.35
- L-threonine
- 44.42~66.63
- L-tryptophan
- 7.808~11.712
- L-tyrosine
- 33.888~50.832
- L-valine
- 43.86~65.79
- Glutathione
- 0.2~0.3
- Para-aminobenzoic acid
- 0.2~0.3
- Biotin
- 0.04148~0.06222
- Calcium pantothenate
- 1.746~2.619
- Choline chloride
- 4.992~7.488
- Folic acid
- 2.06~3.09
- Inositol
- 13.48~20.22
- Niacinamide
- 1.8074~2.7111
- Pyridoxal HCl
- 1.6~2.4
- Pyridoxine HCl
- 0.2124~0.3186
- Riboflavin
- 0.2076~0.3114
- Thiamine HCl
- 1.868~2.802
- Vitamin B12
- 0.273~0.4095
- Hypoxanthine
- 0.816~1.224
- Linoleic acid
- 0.0168~0.0252
- Lipoic acid (thioctic acid)
- 0.042~0.063
- Putrecine dihydrochloride
- 0.0322~0.0483
- Thymidine
- 0.146~0.219
- Sodium chloride
- 5279.8~7919.7
- Potassium chloride
- 284.72~427.08
- Calcium chloride
- 86.644~129.966
- (anhydrous)
- Calcium nitrate 4H2O
- 20~30
- Magnesium chloride
- 11.444~17.166
- (anhydrous)
- Magnesium sulfate
- 48.844~73.266
- (anhydrous)
- Sodium dihydrogen
- 43.48~65.22
- phosphate (anhydrous)
- Disodium monohydrogen
- 188.408~282.612
- phosphate (anhydrous)
- Glucose (anhydrous)
- 1860.4~2790.6
- Ferric nitrate 9H2O
- 0.04~0.06
- Copper sulfate 5H2O
- 0.0005~0.00075
- Ferrous sulfate 7H2O
- 0.1668~0.2502
- Zinc sulfate 7H2O
- 0.1728~0.2592
- Phenol red
- 5.248~7.872.
117 more elements
Dependencies are inferred from the claim text.