Organotypic slice cultures of human gastric and esophagogastric junction cancer
Organotypic slice cultures of human gastric and esophagogastric junction cancer
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DOI：
10.1002/cam4.720
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发表时间：
2016-07-01
期刊：
影响因子：
4
通讯作者：
Lordick, Florian
中科院分区：
文献类型：
--
作者：
Koerfer, Justus;Kallendrusch, Sonja;Lordick, Florian
胃和食道胃交界癌是一种异质性和侵袭性肿瘤，对细胞毒治疗的反应是不可预测的。需要新的方法来分析耐药性。在这里，我们描述了一种新的技术，通过这种技术，人类肿瘤标本可以在体外培养，保留部分自然癌症微环境。用组织切割机将新鲜的手术组织标本切成400亩m的切片，在6孔板中培养长达6天。切片进行常规组织病理学和免疫组织化学检查。细胞角蛋白染色(CK8、AE1/3)检测肿瘤细胞密度，Ki-67染色检测细胞增殖，裂解caspase-3染色检测细胞凋亡。在体外暴露于5-FU和顺铂2-4天后，在幼稚条件下对切片进行分析。切片培养技术可以很好地保存培养期间的组织形态和肿瘤细胞的完整性。化疗后，肿瘤细胞减少，细胞凋亡率增加。可评估肿瘤的药物敏感性。成功地建立了胃和食管胃交界癌的器官型切片培养方法。细胞毒性药物的作用可以被监测。它们可用于研究人体组织中的耐药机制，并可为预测治疗反应提供一个独特而强大的体外平台。
Gastric and esophagogastric junction cancers are heterogeneous and aggressive tumors with an unpredictable response to cytotoxic treatment. New methods allowing for the analysis of drug resistance are needed. Here, we describe a novel technique by which human tumor specimens can be cultured ex vivo, preserving parts of the natural cancer microenvironment. Using a tissue chopper, fresh surgical tissue samples were cut in 400 mu m slices and cultivated in 6-well plates for up to 6 days. The slices were processed for routine histopathology and immunohistochemistry. Cytokeratin stains (CK8, AE1/3) were applied for determining tumor cellularity, Ki-67 for proliferation, and cleaved caspase-3 staining for apoptosis. The slices were analyzed under naive conditions and following 2-4 days in vitro exposure to 5-FU and cisplatin. The slice culture technology allowed for a good preservation of tissue morphology and tumor cell integrity during the culture period. After chemotherapy exposure, a loss of tumor cellularity and an increase in apoptosis were observed. Drug sensitivity of the tumors could be assessed. Organotypic slice cultures of gastric and esophagogastric junction cancers were successfully established. Cytotoxic drug effects could be monitored. They may be used to examine mechanisms of drug resistance in human tissue and may provide a unique and powerful ex vivo platform for the prediction of treatment response.