腹水来源胃癌原代细胞的建立及鉴定  

Primary cell culture and identification of ascites-derived gastric cancer cells

作　　者：姜文静[1] 张忠献[2] 王尧河[2] 石海亮[2] 赵瑞华[1] 宗红[1] 曹风雨[2] 

机构地区：[1]郑州大学第一附属医院肿瘤科,郑州450052 [2]郑州大学中英分子肿瘤学研究中心,郑州450052

出　　处：《郑州大学学报（医学版）》2016年第4期498-502,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金面上项目81272525;郑州市科学技术局科技创新团队项目131PCXTD633;河南省医学科技攻关计划项目201202001

摘　　要：目的:以胃癌患者腹水来源的胃癌细胞为研究对象,旨在探索出一种相对成熟的肿瘤细胞原代培养技术及鉴定方法。方法:对腹水来源胃癌细胞进行原代培养,通过低浓度胰蛋白酶消化法及差速贴壁法进行原代细胞纯化,获得能够稳定传代的胃癌转移瘤细胞系,并通过观察细胞形态、免疫荧光检测、绘制细胞增殖曲线、构建裸鼠皮下移植瘤模型等方法对其进行鉴定。结果:获得1例能够稳定传代的腹水来源胃腺癌细胞株(命名为CFJ),细胞形态一致,表达上皮性生物标志物,细胞核呈现明显的异型性,能在裸鼠皮下成功构建移植瘤模型,增殖和转移能力强,可见裸鼠纵隔、腋窝等多处淋巴结转移。结论:胃癌转移性腹水可以成为建立原代肿瘤细胞的很好来源。Aim： To develop an optimal system for primary cell culture and identification technique with the ascitesderived gastric cancer cells as the research object. Methods： The ascites-derived gastric cancer cells were cultured and purified through the combination of digestion with low concentration trypsin and differential attachment. And the cell morphology was observed by HE and immunocytochemical staining. The cellular proliferation was detected. The tumor growth of established cell line in vivo was also investigated by the subcutaneous transplantation in nude mice model. Results： An ascites-derived gastric adenocarcinoma cell line（ named as CFJ） that could passage steadily had been successfully developed.The cells were consistent in morphology with obvious atypia nuclei and with cytoplasm expressing epithelial biomarkers.Furthermore,it succeeded in establishing the subcutaneous transplantation model in nude mice,and found the cells could locally metastasize to the lymph nodes of mediastinum and axilla in the mice. Conclusion： Metastatic ascites of gastric cancer patients could be a good source of the primary cell culture.

关 键 词：腹水 胃癌 原代培养 转化 成瘤 

分 类 号：R733[医药卫生—肿瘤]