小鼠肝癌原位移植性肿瘤动物模型的改良  

Improved mouse model of orthotopic transplantation for hepatocellular carcinoma

作　　者：罗晓琴 丁冠茗 郑旭 金京燕 刘阳 张楠 王靖宇 王爱国 LUO Xiaoqin;DING Guanming;ZHENG Xu;JIN Jingyan;LIU Yang;ZHANG Nan;WANG Jingyu;WANG Aiguo(Laboratory Animal Center of Dalian Medical University,Dalian 116044,China)

机构地区：[1]大连医科大学实验动物中心,辽宁大连116044

出　　处：《中国比较医学杂志》2021年第6期16-22,共7页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金项目(30872950)。

摘　　要：目的探索建立一套高效、稳定的肝原位移植性肿瘤动物模型的技术方法。方法采用10μL微量注射针将5μL含有5×105个H22细胞溶液沿肝叶长轴种植于小鼠肝左叶,在退出针头避免H22细胞向外溢漏的关键步骤上,分别采用了棉压和热封两种方法,接种后在第19天检测肝肿瘤的大小、重量及病理学变化,比较这两种方法在制备稳定、均一肝原位移植瘤中的效果差异。结果两种方法的肝肿瘤形成率均为100%。两组小鼠的肿瘤大小、重量及病理表现上虽无统计学差异,但与棉压组相比较,热封组肝移植瘤的大小均一,肿瘤重量的波动性小。特别是,在剖检时发现,棉压组出现了高比率的腹水(36.4%)和腹壁瘤(36.4%),而热封组没有出现上述现象,两组统计学差异显著(P<0.05)。说明棉压组的肿瘤细胞从注射孔溢漏的情况显著增高,这也是棉压组肝移植瘤大小波动性较大的根本原因。另外,在棉压组还发现1只发生腹水的小鼠同时也出现肝内多个肿瘤结节,这可能是腹水瘤细胞通过脉管系统形成的肝转移现象。结论本研究成功摸索出一套制备均一、稳定肝原位移植性肿瘤的技术方法和实施要点,具有时间短,易操作,模型稳定等特点,为相关的科研人员提供了良好的借鉴。Objective To establish a high-efficiency and stable mouse model of liver orthotopic transplantation for hepatocellular carcinoma.Methods A 5μL solution containing 5×105 H22 cells was injected into the left liver lobe using a 10μL micro-syringe.In the key step of sealing the pinholes on the liver surface,two method were used to prevent cell leakage:cotton-pressed and heat-sealed.The size,weight,and pathology of liver tumors were examined on the 19th day after inoculation in preparation for establishing a stable and uniform liver orthotopic transplantation model.Results The liver tumor formation rate of both method was 100%.Although there was no statistically significant difference in tumor size,weight,or pathological performance between the two groups,the size and weight of the transplanted liver tumors were more uniform and showed smaller fluctuation in the heat-sealed than cotton-pressed group.In particular,significantly high rates of ascites(36.4%)and abdominal tumors(36.4%)were found in the cotton-pressed group(P<0.05).However,these abnormalities did not develop in the heat-sealed group.This finding indicates that the tumor cells more easily leaked from the pinholes in the cotton-pressed group,which may have been the main cause of the larger fluctuations in the size and weight of orthotopic transplanted liver tumors.In addition,one mouse with multiple abdominal tumors in the cotton-pressed group also exhibited tumor nodules,which may have been the result of metastasis from abdominal or ascitic tumor cells through the vasculature.Conclusions In the present study,we established a method for preparing a uniform and stable mouse model of liver orthotopic transplantation for hepatocellular carcinoma and described the key implementation points.The merits of short time,an easy operation,and stability provide a good reference for liver cancer research.

关 键 词：肝癌 H22 肝原位移植 热封 

分 类 号：R-33[医药卫生]