Hamilton W G, Ham R G
In Vitro. 1977 Sep;13(9):537-47. doi: 10.1007/BF02627849.
A protein-free synthetic medium, MCDB 301, has been developed for clonal growth of Chinese hamster ovary cell lines. Medium F12 was developed originally for that purpose, but later failed to support good growth without small amounts of serum protein. Growth was restored by the addition of nonphysiological amounts of commercially prepared thyroxine or smaller amounts of the trace element selenium. The thyroxine preparation was shown to contain sufficient selenium to account for all of its growth-promoting activity. MCDB 301 contains increased concentrations of calcium chloride and glutamine, and a smaller amount of cysteine than medium F12. It also has been supplemented with 19 inorganic ions, in addition to selenium and those in medium F12, in order to insure against possible future deficiencies as chemicals are purified further. A Chinese hamster lung line which will not grow in MCDB 301 alone will grow when the medium is supplemented either with methylcellulose or with insulin. The growth-promoting activity is thought to be an impurity shared in common by both substances. The probable "essential" role of impurities in cellular growth in most synthetic media and the problems involved in attempting to develop a truly "defined" medium are discussed.
一种无蛋白合成培养基MCDB 301已被开发用于中国仓鼠卵巢细胞系的克隆生长。培养基F12最初是为此目的而开发的，但后来发现如果没有少量血清蛋白，就无法支持良好的生长。通过添加非生理量的市售甲状腺素或少量的微量元素硒可恢复生长。已证明甲状腺素制剂中含有的硒足以解释其所有的促生长活性。MCDB 301中氯化钙和谷氨酰胺的浓度增加，半胱氨酸的量比培养基F12少。除了硒和培养基F12中的那些离子外，它还添加了19种无机离子，以确保随着化学物质进一步纯化，未来不会出现可能的缺乏。一种单独在MCDB 301中不能生长的中国仓鼠肺细胞系，当培养基添加甲基纤维素或胰岛素时就会生长。据认为，促进生长的活性是这两种物质共有的一种杂质。讨论了大多数合成培养基中杂质在细胞生长中可能的“必需”作用以及试图开发真正“限定”培养基所涉及的问题。
