Kaighn M E, Kumari H L, Spalding J W, Barrett J C, Iype P T
Biological Research Faculty and Facility, Inc., Ijamsville, MD 21754.
Carcinogenesis. 1993 Apr;14(4):579-84. doi: 10.1093/carcin/14.4.579.
The objective of this work was to develop a serum-free medium that supported the growth of secondary Syrian hamster embryo (SHE) cells suitable for use in the morphological transformation assay (MTA). The current assay employs high levels of fetal bovine serum, each lot of which must be selected for its efficacy in the assay. To circumvent the problems associated with the use of serum, two serum-free media (CGM-4 and CGM-9) were developed. The approach used was to replace serum with more defined hormones and growth factors. DME/F12 with low NaHCO3 (pH 6.7) was chosen as a basal nutrient medium since this pH has been reported to give enhanced transformation. Both serum-free media supported the clonal high density growth of normal secondary SHE cells. These media were also found to support the growth of several established SHE cell lines exhibiting successive stages of neoplastic progression. However, significant differential responses of the individual cell lines to CGM-4 and CGM-9 were observed, whereas, in serum-supplemented medium all three lines responded similarly. These results raise the hope that CGM-4 and CGM-9 will be useful for studies of altered autocrine function during transformation and progression as well as for use in the transformation assay itself.
这项工作的目的是开发一种无血清培养基,以支持适合用于形态转化试验(MTA)的二代叙利亚仓鼠胚胎(SHE)细胞的生长。当前的试验使用高浓度的胎牛血清,每一批血清都必须根据其在试验中的效力进行选择。为了规避与使用血清相关的问题,开发了两种无血清培养基(CGM - 4和CGM - 9)。所采用的方法是用更明确的激素和生长因子替代血清。选择低碳酸氢钠(pH 6.7)的DME/F12作为基础营养培养基,因为据报道这个pH值能增强转化。两种无血清培养基都支持正常二代SHE细胞的克隆高密度生长。还发现这些培养基能支持几种已建立的处于肿瘤进展连续阶段的SHE细胞系的生长。然而,观察到各个细胞系对CGM - 4和CGM - 9有显著的差异反应,而在补充血清的培养基中,所有三个细胞系的反应相似。这些结果让人希望CGM - 4和CGM - 9将有助于研究转化和进展过程中自分泌功能的改变,以及用于转化试验本身。