Ultrastructural and immunocytochemical characterization of an immortalized human breast epithelial cell line, MCF-10.
Ultrastructural and immunocytochemical characterization of an immortalized human breast epithelial cell line, MCF-10.
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发表时间：
1990-09
期刊：
影响因子：
11.2
通讯作者：
L. Tait;H. Soule;J. Russo
中科院分区：
文献类型：
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作者：
L. Tait;H. Soule;J. Russo
人乳腺上皮细胞系MCF-10来源于一名患有纤维囊性疾病的36岁、已生育、绝经前妇女的s.c.乳房切除术组织。MCF-10M细胞在1.05 mM钙溶液中连续转移后开始衰老。这些细胞自发地产生了两个不朽的子细胞，MCF-10A，或附着细胞，和MCF-10F，或漂浮细胞，它们在Dulbecco改良的基本培养基和Ham的F-12中增殖了4年多，要么是通常的钙浓度为1.05 mM (DMEM-H)，要么是在含有0.04 mM钙或低钙的培养基中。本文报道的研究表明MCF-10是乳腺上皮细胞系。电镜显示MCF-10A和MCF-10F均具有管腔导管细胞的特征，而不具有肌上皮细胞的特征。当分别在Dulbecco改良的基本培养基- ham 's F-12和低钙培养基中培养1200多天后，它们保持了上皮特征，尽管钙浓度对细胞形态有很强的影响。在Dulbecco's改良Eagle's培养基中培养的细胞是低立方的，有大量的桥粒和较短的微绒毛，而在低钙培养基中培养的细胞比在DMEM-H中培养的细胞有明显减少的桥粒数量，更球形，有更多的微绒毛，且微绒毛更长。通过免疫细胞化学检测上皮唾液黏液蛋白和角蛋白，证实了这些细胞的乳腺上皮起源。采用单克隆抗体MFA-breast、MC5和多克隆抗体上皮膜抗原检测上皮性唾液黏液蛋白。用KA-4、K-14、AE1/AE3和K-19特异性抗体对角蛋白进行表征。结论：MCF-10A和MCF-10F细胞是乳腺上皮细胞，是研究生长和癌变基本过程的重要工具。
The human breast epithelial cell line MCF-10 was derived from s.c. mastectomy tissue from a 36-year-old, parous, premenopausal woman with fibrocystic disease. It was initiated as a mortal cell line (MCF-10M), which senesces when transferred serially in 1.05 mM calcium. These cells spontaneously gave origin to two immortal sublines, MCF-10A, or attached cells, and MCF-10F, or floating cells, which have proliferated for more than 4 years in Dulbecco's modified essential medium and Ham's F-12 either with the customary calcium concentration of 1.05 mM (DMEM-H) or in medium containing 0.04 mM calcium or low calcium. Studies reported here indicate that MCF-10 is a mammary epithelial cell line. Electron microscopy showed that both MCF-10A and MCF-10F have characteristics of luminal ductal cells, but not of myoepithelial cells. When grown for more than 1200 days in Dulbecco's modified essential medium-Ham's F-12 and low calcium media, respectively, they maintained their epithelial characteristics, although the concentration of calcium exerted a powerful influence on cell morphology. Cells grown in Dulbecco's modified Eagle's medium-Ham's F12 medium are low cuboidal with numerous desmosomes and short microvilli, whereas those grown in low calcium medium have significantly reduced number of desmosomes, are more spherical, and have greater numbers of microvilli which are longer than those of cells grown in DMEM-H. The breast epithelial orgin of these cells was confirmed by immunocytochemical detection of epithelial sialomucins and keratins. The monoclonal antibodies MFA-breast and MC5 and the polyclonal antibody epithelial membrane antigen were used to detect the epithelial sialomucins. Keratins were characterized by using KA-4, K-14, AE1/AE3, and K-19 specific antibodies. It was concluded that MCF-10A and MCF-10F cells are breast epithelial cells and that they represent an important tool for studies of the basic processes of growth and carcinogenesis.