Shipley G D, Ham R G
In Vitro. 1981 Aug;17(8):656-70. doi: 10.1007/BF02628401.
Improved culture conditions have been developed that will support clonal growth of Swiss mouse embryo 3T3 cells at concentrations of serum protein at low as 125 micrograms/ml. Survival of the cells under completely protein-free conditions also is enhanced greatly. The improvements that made these results possible include: (a) use of medium MCDB 402, which was developed specifically for Swiss 3T3 cells by adjusting the concentrations of all components of Dulbecco's modified Eagle's medium to optimum values for clonal growth with minimal serum protein and by adding other nutrients such as trace elements and "nonessential" amino acids that were not in the original formula; (b) use of culture surfaces that are coated with a positively charged polymer, poly-D-lysine; and (c) use of gentle low temperature trypsinization technique that minimizes cellular damage and the need to neutralize residual trypsin.
已经开发出了改良的培养条件，这种条件能够支持瑞士小鼠胚胎3T3细胞在血清蛋白浓度低至125微克/毫升的情况下进行克隆生长。在完全无蛋白的条件下，细胞的存活率也大大提高。促成这些结果的改进包括：(a) 使用MCDB 402培养基，该培养基是专门为瑞士3T3细胞开发的，通过将杜尔贝科改良伊格尔培养基的所有成分浓度调整到以最低血清蛋白进行克隆生长的最佳值，并添加其他营养物质如微量元素和原始配方中没有的“非必需”氨基酸；(b) 使用涂有带正电荷聚合物聚-D-赖氨酸的培养表面；(c) 使用温和的低温胰蛋白酶消化技术，该技术可将细胞损伤降至最低，并减少中和残留胰蛋白酶的需求。
