Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and <em>Ex vivo</em> Fibroblasts

Marc D. Bullock; Max Mellone; Karen M. Pickard; Abdulkadir Emre Sayan; Richard Mitter; John N. Primrose; Graham K. Packham; Gareth Thomas; Alexander H. Mirnezami

doi:10.3791/51475

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Medicine

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JoVE Journal Medicine

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

在结直肠癌细胞的三维模型的入侵肿瘤微环境的分子剖析和体外成纤维细胞

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10:33 min

April 29, 2014

DOI: 10.3791/51475-v

Marc D. Bullock^1,2, Max Mellone¹, Karen M. Pickard¹, Abdulkadir Emre Sayan¹, Richard Mitter³, John N. Primrose², Graham K. Packham¹, Gareth Thomas¹, Alexander H. Mirnezami^1,2

¹Cancer Sciences Unit,University of Southampton School of Medicine, ²University Surgical Unit,University of Southampton School of Medicine, ³Bioinformatics Unit,London Research Institute, Cancer Research UK

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激光显微切割细胞和基质由合成的，组织工程肠癌模型的分子分析代表了一种新的和可操作的方法在肿瘤细胞之间的界面来表征和解剖的独特生物学在浸润前沿和癌症相关的基质细胞。

该程序的总体目标是表征侵袭性肿瘤前部恶性上皮细胞与癌症相关基质细胞之间界面的独特生物学特征。这是通过首先从结肠植入物建立原代成纤维细胞培养物来实现的。第二步是使用原代结肠成纤维细胞和培养的结直肠癌细胞构建器官型共培养模型。

接下来，将器官型模型固定在福尔米蛋白中，切片到膜安装的载玻片上并用牙槽槽嵴紫染色。最后一步是激光显微切割，以便将侵袭的肿瘤细胞与非侵袭的肿瘤细胞和癌症相关的基质细胞分开。最终，分子分析技术可用于表征周围肿瘤微环境中侵袭和非侵袭肿瘤细胞和基质细胞的生物学特性。

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医药第86期大肠癌癌症转移器官培养激光显微切割分子分析浸润肿瘤微环境间质组织上皮细胞成纤维细胞