Short-term primary culture of epithelial cells derived from human breast tumours
Short-term primary culture of epithelial cells derived from human breast tumours
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DOI：
10.1038/bjc.1998.702
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发表时间：
1998-12-01
影响因子：
8.8
通讯作者：
Atkin, SL
中科院分区：
文献类型：
--
作者：
Speirs, V;Green, AR;Atkin, SL
作为乳腺癌的实验模型，大多数研究依赖于已建立的人类乳腺癌细胞系。然而，其中许多细胞系是 20 多年前建立的，其中许多来自胸腔积液而不是原发肿瘤，因此使用它们作为代表性模型的有效性值得怀疑。本文描述了我们在三年多的时间里，通过差速离心和选择性培养基培养，从原发性乳腺肿瘤中建立短期上皮细胞富集制剂的经验。 55% 的样本中成功培养了上皮细胞，但培养成功似乎与肿瘤组织学、分期、分级或淋巴结状态无关。富含上皮细胞的培养物对广谱细胞角蛋白和上皮膜抗原（EMA）呈免疫阳性。角蛋白19的阳性证实了培养物含有肿瘤来源的细胞，其还显示出I型类固醇转化酶17β-羟基类固醇脱氢酶的还原途径的显着更高的活性。通过肿瘤来源的培养物中完全不存在钙调蛋白样基因NB-1进一步证实了培养物含有肿瘤而不是正常上皮细胞；这仅与正常乳腺上皮有关。从雌激素受体 (ER) 阳性肿瘤建立的培养物中，85% 在体外表达 ER；尽管 ER 阳性表型随着时间的推移逐渐消失，但这一功能在 66% 的培养物中发挥了作用。总之，无论组织病理学或临床细节如何，上皮细胞都可以从原发性乳腺肿瘤中分离并维持为短期培养物，从而提供了比乳腺癌细胞系具有更大生物学和临床相关性的模型系统。
As experimental models for breast cancer, most studies rely on established human breast cancer cell lines. However, many of these lines were established over 20 years ago, many from pleural effusions rather than the primary tumour, so the validity of using them as representative models is questionable. This paper describes our experiences, over a 3-year period, in establishing short-term epithelial-cell-enriched preparations from primary breast tumours based on differential centrifugation followed by culture in selective media. Epithelial cells were successfully cultured from 55% of samples, but culture success did not appear to be correlated with tumour histology, stage, grade or node status. Epithelial cell-enriched cultures were immunopositive for broad-spectrum cytokeratin and epithelial membrane antigen (EMA). Positivity for keratin 19 confirmed that the cultures contained tumour-derived cells, which additionally showed significantly higher activity of the reductive pathway of the steroid-converting enzyme 17 beta-hydroxysteroid dehydrogenase type I. That the cultures contained tumour and not normal epithelial cells was further substantiated by the complete absence of the calmodulin-like gene NB-l in tumour-derived cultures; this is only associated with normal breast epithelia. Eighty-five per cent of cultures established from oestrogen receptor (ER)-positive tumours expressed ER in vitro; this was functional in 66% of cultures, although ER-positive phenotype was gradually lost over time. In conclusion, epithelial cells can be isolated and maintained as short-term cultures from primary breast tumours irrespective of histopathological or clinical details, providing a model system with a greater biological and clinical relevance than breast cancer cell lines.