Bertheussen K
Institute of Medical Biology, University of Tromsø, Norway.
Cytotechnology. 1993;11(3):219-31. doi: 10.1007/BF00749873.
The development of a new stable synthetic serum replacement (SSR) is described, which allows the cultivation of mammalian cells in a defined, protein-free medium containing only dialyzable components. With a low concentration of insulin (RPMI-SR2 medium), growth rates of the transformed cell lines L929, HELA S3, and the hybridoma 1E6 were comparable to growth rates obtained with a serum-containing medium. The same medium also supported long-term cultivation of non-dividing mouse macrophages. The main principle of SSR is a metal ion buffer containing a balanced mixture of iron and trace metals. Stability against precipitation of important metals is achieved by the combined use of EDTA and citric acid as chelating agents. Efficient iron supply is mediated through the inclusion of the compound Aurintricarboxylic acid as a synthetic replacement for transferrin. SSR also contains a growth-promoting surfactant, Pluronic F68. Thus SSR provides a general foundation for growth and differentiation normally provided by serum. Limitations of other serum-free medium designs are discussed here: 1) the inability of transferrin to chelate all metals in the medium; and 2) the use of inorganic iron salts or iron citrate as an iron supplement leads to rapid precipitation of iron hydroxide in the medium. Both these problems are solved in the design of SSR.
本文描述了一种新型稳定合成血清替代品（SSR）的开发，它能够使哺乳动物细胞在仅含可透析成分的特定无蛋白培养基中培养。在低浓度胰岛素（RPMI - SR2培养基）的情况下，转化细胞系L929、HELA S3以及杂交瘤1E6的生长速率与在含血清培养基中获得的生长速率相当。同样的培养基也支持非分裂小鼠巨噬细胞的长期培养。SSR的主要原理是一种含有铁和微量金属平衡混合物的金属离子缓冲液。通过联合使用EDTA和柠檬酸作为螯合剂，实现了重要金属抗沉淀的稳定性。通过加入金精三羧酸作为转铁蛋白的合成替代品来介导有效的铁供应。SSR还含有一种促进生长的表面活性剂，普朗尼克F68。因此，SSR为通常由血清提供的生长和分化提供了一个通用基础。本文讨论了其他无血清培养基设计的局限性：1）转铁蛋白无法螯合培养基中的所有金属；2）使用无机铁盐或柠檬酸铁作为铁补充剂会导致培养基中铁氢氧化物迅速沉淀。在SSR的设计中解决了这两个问题。
