作者：胡蕾1，陈伟1，张世维2，蒋菲1，文璋1，刘宇虹3，胡臻然2，曹清华4，杨峥1

单位：1.中山大学附属第七医院 病理科，广东 深圳 518000；2.中山大学附属第七医院 消化医学中心，广东 深圳 518000；3.中山大学附属第七医院 普外科，广东 深圳 518000；4.中山大学附属第一医院 病理科，广东 广州 510080

Authors: Hu Lei1, Chen Wei1, Zhang Shiwei2, Jiang Fei1, Wen Zhang1, Liu Yuhong3, Hu Zhenran2, Cao Qinghua4,Yang Zheng1

Unit: 1.Department of Pathology, The Seventh Affiliated Hospital of Sun Yat -Sen University, Shenzhen 518000,Guangdong,China；2.Digestive Diseases Center, The Seventh Affiliated Hospital of Sun Yat -Sen University, Shenzhen 518000,Guangdong, China；3.General Surgery, The Seventh Affiliated Hospital of Sun Yat -Sen University, Shenzhen 518000,Guangdong,China；4.Department of Pathology, The First Affiliated Hospital of Sun Yat -Sen University, Guangzhou 510080,Guangdong,China

摘要：

目的 建立胃癌恶性腹水来源类器官模型,并将该模型用于化疗药物筛查。方法 收集4例胃癌患者的腹水,离心处理后与基质胶混合种板,然后添加含有生长因子的培养基进行类器官培养。采用苏木精-伊红染色和免疫组织化学染色法对胃癌腹水来源类器官和原始恶性腹水肿瘤细胞进行病理形态学及相关免疫标志物分析。通过对2种化疗药物顺铂和氟尿嘧啶的敏感性筛查,评估胃癌恶性腹水来源类器官用于药物筛选的适用性。结果 成功培养3例胃癌恶性腹水来源类器官,类器官的形态和生长特征存在个体差异性,所有类器官均可稳定传代、冻存与复苏。胃癌恶性腹水来源类器官保留了相应恶性腹水肿瘤细胞的组织结构特征,相关免疫标志物表达基本保持一致。3例类器官对2种化疗药物表现出不同的药物敏感性,对氟尿嘧啶均耐药,对顺铂的耐药存在个体差异。结论 本研究成功建立胃癌恶性腹水来源类器官模型,该模型很好地保持了胃癌恶性腹水原始肿瘤细胞的病理学特征。类器官对2种化疗药物的反应初步证实其作为体外药物筛选模型的可行性。

关键词：胃癌恶性腹水; 类器官; 药物敏感性筛查; 个性化医疗

Abstract：

Objective The purpose of this study was to establish a gastric cancer-derived organoid model from malignant ascites and to use this model for chemotherapeutic drug screening. Method The ascites of 4 patients with gastric cancer were collected, they were mixed with Matrigel and seeded after centrifugation. The medium containing growth factors was added for organoid culture. Hematoxylin -eosin staining and immunohistochemical staining were used to analyze the pathological morphology and related immune markers of gastric cancer ascites -derived organoids and primitive malignant ascites tumor cells. The applicability of gastric cancer-derived organoids from malignant ascites for drug screening was assessed by sensitivity screening to two chemotherapeutic drugs include cisplatin and fluorouracil. Result Three cases of gastric cancer -derived organoids from malignant ascites were successfully cultured. There were individual differences in the morphology and growth characteristics of the organoids. All organoids could be stably passaged, cryopreserved, and resuscitated. The malignant ascites -derived organoids of gastric cancer retained the histological characteristics of the corresponding malignant ascites tumor cells, and the expression of related immune markers remained basically the same. The three organoids showed different drug sensitivities to the two chemotherapeutic drugs, all were resistant to fluorouracil, and there were individual differences in the resistance to cisplatin. Conclusion In this study, an organoid model derived from malignant ascites of gastric cancer was successfully established, which maintained the pathological characteristics of the original tumor cells of malignant ascites of gastric cancer well. The organoid response to two chemotherapeutic drugs preliminarily confirmed its feasibility as an in vitro drug screening model.

Key Words: Malignant ascites of gastric cancer; Organoids; Drug screening; Personalized medicine