Capturing complex tumour biology in vitro: histological and molecular characterisation of precision cut slices.
Capturing complex tumour biology in vitro: histological and molecular characterisation of precision cut slices.
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DOI：
10.1038/srep17187
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发表时间：
2015-12-09
影响因子：
4.6
通讯作者：
Hickman JA
中科院分区：
文献类型：
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作者：
Davies EJ;Dong M;Gutekunst M;Närhi K;van Zoggel HJ;Blom S;Nagaraj A;Metsalu T;Oswald E;Erkens-Schulze S;Delgado San Martin JA;Turkki R;Wedge SR;af Hällström TM;Schueler J;van Weerden WM;Verschuren EW;Barry ST;van der Kuip H;Hickman JA
体内肿瘤的精确切割切片允许在体外询问来自实体瘤的异质细胞及其天然微环境。它们提供了低通量但高含量的体外实验平台。使用小鼠模型作为三种常见的人类实体瘤的替代物，我们描述了一个标准化的工作流程，用于系统比较肿瘤切片培养方法和基于组织微阵列的方法来存档它们。使用免疫组织化学和转录生物标志物，特别是细胞应激，将培养的切片与其体内来源组织进行比较。机械切片引起最小应力。肿瘤切片的培养需要器官型支持材料和大气氧以维持完整性，并且与蛋白质表达的显著时间和局部变化相关，例如HIF-1α。我们建议坚持所描述的强大的工作流程，在通过药理学或其他手段询问肿瘤切片之前识别蛋白质表达的时空变化。
Precision-cut slices of in vivo tumours permit interrogation in vitro of heterogeneous cells from solid tumours together with their native microenvironment. They offer a low throughput but high content in vitro experimental platform. Using mouse models as surrogates for three common human solid tumours, we describe a standardised workflow for systematic comparison of tumour slice cultivation methods and a tissue microarray-based method to archive them. Cultivated slices were compared to their in vivo source tissue using immunohistochemical and transcriptional biomarkers, particularly of cellular stress. Mechanical slicing induced minimal stress. Cultivation of tumour slices required organotypic support materials and atmospheric oxygen for maintenance of integrity and was associated with significant temporal and loco-regional changes in protein expression, for example HIF-1α. We recommend adherence to the robust workflow described, with recognition of temporal-spatial changes in protein expression before interrogation of tumour slices by pharmacological or other means.