SERUM-FREE MAMMALIAN CELL CULTURE MEDIUM, AND USES THEREOF

摘要

Mammalian cells are cultured, in vitro, suspended in a serum-free medium (A) containing >= 1 polyanionic or polycationic compound (I). Also new are: (1) (A); (2) virus produced in mammalian cells cultured in (A); (3) polypeptide produced in genetically engineered mammalian cells cultured in (A); (4) culture medium (A') for mammalian cells containing a ferrous chelate and zinc salt; (5) improved medium (A'') for growing Chinese hamster ovary (CHO) cells in which insulin is replaced by Zn salt and/or transferrin by a ferrous and/or ferric chelate. (I) is a polysulphated or polysulphonated compound, preferably dextran, heparan, chondroitin, dermatan, pentosan sulphates, heparin or a proteoglycan, best dextran sulphate (DS) of molecular weight about 5000. (A) is free of protein and includes amino acids, vitamins, inorganic salts, buffers, sugars, lipids, transferrin and insulin (or alternatives to these two), optionally also cytokines, heparin or peptides, particularly from rice or soya. A preferred basal medium contains ethanolamine; glucose; Hepes (N-(2-hydroxyethyl)- piperazine-N'-(2-ethanesulphonic acid)); insulin; transferrin; linolenic acid; lipoic acid; phenol red; etc. (A') contains 0.28-110 (preferably about 11) mg/l ferrous ions and 0.07-7.3 (preferably about 0.354) mg/l Zn ions, with similar levels in (A''). It may also include (I) and other components as for the basal medium described. (I) is present at up to 500 (preferably about 100) mg/l. The media have pH 7-7.6 (especially 7.2-7.4).