Application of HEPES|CAS 7365-45-9| in cell culture medium

HEPES is a zwitterionic buffer, belonging to good's buffer, with pKa 7.45 at 25°C and pH buffer range of 6.8-8.2, so it is widely used in cell culture media, IVD kits, cosmetics, medicine and other fields. CN102827804A patent introduces the application of HEPES in cell culture medium. The medium mainly consists of 7 parts, specifically: vitamins, lipids, amino acids, inorganic salts, protein hydrolyzates, buffer components and additives. Details as follows:

1. Vitamins consist of 12 ingredients, specifically inositol, vitamin C, pyridoxine phosphate, pyridoxal phosphate, cobalamin, nicotinamide, riboflavin, thiamine hydrochloride, calcium D-pantothenate, folic acid, Biotin and Choline Chloride;

2. Lipids are composed of 2 components, specifically linoleic acid and PluronicF-68; 

3. Amino acids are composed of 21 components, specifically L-tryptophan, L-cysteine, L-isoleucine, L-leucine, L-valine, L-arginine hydrochloride Salt, L-Histidine Hydrochloride, L-Lysine, L-Methionine, L-Glutamic Acid, L-Phenylalanine, L-Aspartic Acid, Glycine, L-Proline, L- Alanine, L-aspartic acid, L-serine, L-threonine, L-glutamine, L-cystine hydrochloride and L-tyrosine disodium salt;

4. Inorganic salt is composed of 11 components, specifically magnesium chloride hexahydrate, magnesium sulfate anhydrous, calcium chloride anhydrous, potassium chloride, sodium chloride, sodium dihydrogen phosphate monohydrate, disodium hydrogen phosphate, heptahydrate sulfuric acid Zinc, ferric nitrate nonahydrate, sodium selenite and copper sulfate pentahydrate;   

5. The protein hydrolyzate can be milk protein hydrolyzate, yeast hydrolyzate or plant hydrolyzate;

6. The buffer components are sodium bicarbonate and HEPES;

7. The additive is composed of 6 components, specifically D-glucose, sodium pyruvate, reduced glutathione, hypoxanthine sodium salt, putrescine and lipoic acid.

This patent has the following beneficial effects:

1. The medium is prepared by conventional methods, which can support the growth of Vero on square bottles and roller bottles, and can support Vero cell microcarrier suspension culture and microcarrier suspension scale-up culture. In addition, it can also be used as a virus maintenance solution to achieve virus serum-free production.

2. The cost is low, and large-scale production can be realized.

References

CN102827804A Medium and method suitable for Vero cell microcarrier suspension amplification culture.