Medina D, Oborn C J, Kittrell F S, Ullrich R L
J Natl Cancer Inst. 1986 Jun;76(6):1143-56.
Immunocytochemical and in vivo transplantation methods were used to study the characteristics of a series of newly developed mammary epithelial cell lines. These mouse mammary cell lines were derived from mid-pregnant primiparous BALB/c female mice and were routinely grown in Dulbecco's modified Eagle medium supplemented with 1% fetal bovine serum, insulin, transferrin, epidermal growth factor, and selenite. Of the 6 cell lines, 1 cell line, COMMA-D, produced normal and preneoplastic mammary outgrowths when it was transplanted into mammary fat pads of syngeneic mice. One cell line, MOD, produced only mammary adenocarcinomas. The other 4 cell lines, COMMA-F, COMMA-T, MOMA-1, and MOMA-2, produced neither normal nor neoplastic outgrowths. Immunocytochemical staining with polyclonal antibodies to keratin and vimentin intermediate filament proteins revealed that 5 of the 6 cell lines were epithelial. The sixth cell line, MOMA-2, was apparently of fibroblastic origin. The COMMA-D cell line was unique compared to the other cell lines with respect to several characteristics. The cell line was morphologically heterogeneous as determined by phase-contrast microscopy, cytologically heterogeneous as determined by immunocytochemical staining, and heterogeneous with respect to DNA content. Finally, the full morphogenic potential of COMMA-D included not only normal mammary ductal and preneoplastic mammary alveolar outgrowths but also adenocarcinomas and fibrosarcomas. The expression of this morphogenic potential upon transplantation in vivo was drastically diminished after passage 14. The significance of the cellular heterogeneity with respect to expression of mammary-specific morphogenesis is not understood at this time; however, conceivably, the observed heterogeneity reflects an essential requirement for morphogenesis in vivo. The transplantation and immunocytochemical characteristics provide the descriptive basis for further studies on these cell lines to determine the cell lineages involved in morphogenesis and preneoplastic transformation in vivo.
采用免疫细胞化学和体内移植方法研究了一系列新建立的乳腺上皮细胞系的特性。这些小鼠乳腺细胞系源自怀孕中期的初产BALB/c雌性小鼠,常规培养于添加1%胎牛血清、胰岛素、转铁蛋白、表皮生长因子和亚硒酸盐的杜尔贝科改良伊格尔培养基中。在这6个细胞系中,1个细胞系COMMA-D移植到同基因小鼠的乳腺脂肪垫中时可产生正常和癌前乳腺增生。1个细胞系MOD仅产生乳腺腺癌。另外4个细胞系COMMA-F、COMMA-T、MOMA-1和MOMA-2既不产生正常增生也不产生肿瘤增生。用针对角蛋白和波形蛋白中间丝蛋白的多克隆抗体进行免疫细胞化学染色显示,6个细胞系中有5个为上皮细胞系。第6个细胞系MOMA-2显然起源于成纤维细胞。COMMA-D细胞系在几个特性方面与其他细胞系相比具有独特性。通过相差显微镜观察,该细胞系在形态上具有异质性;通过免疫细胞化学染色确定,在细胞学上具有异质性;在DNA含量方面也具有异质性。最后,COMMA-D的完全形态发生潜能不仅包括正常乳腺导管和癌前乳腺腺泡增生,还包括腺癌和纤维肉瘤。传代14次后,这种形态发生潜能在体内移植时的表达大幅降低。目前尚不清楚细胞异质性对于乳腺特异性形态发生表达的意义;然而,可以想象,观察到的异质性反映了体内形态发生的基本要求。移植和免疫细胞化学特性为进一步研究这些细胞系提供了描述性基础,以确定体内形态发生和癌前转化所涉及的细胞谱系。