Leeper Alexander D, Farrell Joanne, Dixon J Michael, Wedden Sarah E, Harrison David J, Katz Elad
Breakthrough Breast Cancer Research Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh.
J Vis Exp. 2011 Jul 29(53):3085. doi: 10.3791/3085.
Breast cancer is a leading cause of mortality in the Western world. It is well established that the spread of breast cancer, first locally and later distally, is a major factor in patient prognosis. Experimental systems of breast cancer rely on cell lines usually derived from primary tumours or pleural effusions. Two major obstacles hinder this research: (i) some known sub-types of breast cancers (notably poor prognosis luminal B tumours) are not represented within current line collections; (ii) the influence of the tumour microenvironment is not usually taken into account. We demonstrate a technique to culture primary breast cancer specimens of all sub-types. This is achieved by using three-dimensional (3D) culture system in which small pieces of tumour are embedded in soft rat collagen I cushions. Within 2-3 weeks, the tumour cells spread into the collagen and form various structures similar to those observed in human tumours1. Viable adipocytes, epithelial cells and fibroblasts within the original core were evident on histology. Malignant epithelial cells with squamoid morphology were demonstrated invading into the surrounding collagen. Nuclear pleomorphism was evident within these cells, along with mitotic figures and apoptotic bodies. We have employed Optical Projection Tomography (OPT), a 3D imaging technology, in order to quantify the extent of tumour spread in culture. We have used OPT to measure the bulk volume of the tumour culture, a parameter routinely measured during the neo-adjuvant treatment of breast cancer patients to assess response to drug therapy. Here, we present an opportunity to culture human breast tumours without sub-type bias and quantify the spread of those ex vivo. This method could be used in the future to quantify drug sensitivity in original tumour. This may provide a more predictive model than currently used cell lines.
乳腺癌是西方世界主要的致死原因。乳腺癌首先在局部扩散，随后向远处扩散，这一过程是影响患者预后的主要因素，这一点已得到充分证实。乳腺癌的实验系统通常依赖于源自原发性肿瘤或胸腔积液的细胞系。有两个主要障碍阻碍了这项研究：（i）目前的细胞系库中没有涵盖一些已知的乳腺癌亚型（尤其是预后较差的管腔B型肿瘤）；（ii）通常没有考虑肿瘤微环境的影响。我们展示了一种培养所有亚型原发性乳腺癌标本的技术。这是通过使用三维（3D）培养系统实现的，在该系统中，小块肿瘤被嵌入柔软的大鼠I型胶原垫中。在2至3周内，肿瘤细胞扩散到胶原中，并形成各种类似于在人类肿瘤中观察到的结构。组织学检查显示，原始核心内有存活的脂肪细胞、上皮细胞和成纤维细胞。具有鳞状形态的恶性上皮细胞侵入周围的胶原。这些细胞内可见核多形性，以及有丝分裂象和凋亡小体。我们采用了光学投影断层扫描（OPT）这一3D成像技术，以量化培养物中肿瘤扩散的程度。我们使用OPT测量肿瘤培养物的总体积，这是在乳腺癌患者新辅助治疗期间常规测量的一个参数，用于评估对药物治疗的反应。在这里，我们提供了一个机会，可以无亚型偏向地培养人类乳腺肿瘤，并在体外量化其扩散情况。这种方法未来可用于量化原始肿瘤中的药物敏感性。这可能提供一个比目前使用的细胞系更具预测性的模型。
