Biology Research Division, Wellcome Research Laboratories, Langley Court, Beckenham, Kent, UK.
Cytotechnology. 1995 Oct;17(3):153-63. doi: 10.1007/BF00749653.
A serum-free medium, WCM5, has been developed for the large scale propagation of CHO (Chinese hamster ovary) cells which express recombinant protein using dihydrofolate reductase as a selectable marker. WCM5 was prepared by supplementing Iscoves medium without lecithin, albumin or transferrin with a number of components which were shown to benefit growth. WCM5 medium contained 5 mg l(-1) human recombinant insulin (Nucellin) but was otherwise protein-free. CHO 3D11() cells which had been engineered to express a humanised antibody, CAMPATH()-1H, were routinely grown using serum-containing medium. From a seeding density of 10(5) cells ml(-1), cells grown in static culture with serum reached a maximal cell density of 6.5×10(5) cells ml(-1) after 6 days in culture and produced a maximal antibody concentration of 69 mg l(-1) after 11 days in culture. CHO 3D11(*) cells grown with serum were washed in serum-free medium then cultured in WCM5 medium. Following a period of adaptation the cell growth and product yield was superior to that achieved with serum-containing medium. CHO cells producing CAMPATH-1H grown in an 8000 l stirred bioreactor seeded with 2×10(5) cells ml(-1) reached a maximal viable cell density of 2.16×10(6) cells ml(-1) after 108 h in culture and a maximal antibody concentration of 131.1 mg l(-1) after 122 h in culture.
一种无血清培养基 WCM5 已被开发用于大规模繁殖表达二氢叶酸还原酶作为选择标记的重组蛋白的 CHO（中国仓鼠卵巢）细胞。WCM5 通过向不含卵磷脂、白蛋白或转铁蛋白的 Iscoves 培养基中添加一些对生长有益的成分来制备。WCM5 培养基含有 5mg/L 的人重组胰岛素（Nucellin），但不含蛋白质。CHO 3D11()细胞经过工程改造表达了一种人源化抗体 CAMPATH()-1H，通常在含血清的培养基中生长。从 10(5)个细胞/ml 的接种密度开始，在含血清的静态培养中，细胞在培养 6 天后达到最大细胞密度 6.5×10(5)个细胞/ml，在培养 11 天后达到最大抗体浓度 69mg/L。用血清培养的 CHO 3D11(*)细胞在无血清培养基中洗涤，然后在 WCM5 培养基中培养。经过一段时间的适应，细胞生长和产物产量优于含血清培养基。在接种密度为 2×10(5)个细胞/ml 的 8000L 搅拌生物反应器中培养的表达 CAMPATH-1H 的 CHO 细胞，在培养 108 小时后达到最大活细胞密度 2.16×10(6)个细胞/ml，在培养 122 小时后达到最大抗体浓度 131.1mg/L。
