建立脑肿瘤裸小鼠原位移植模型的方法探讨  

Preliminary study on methods of establishment of brain tumor orthotopic transplantation model in nude mice

作　　者：李如军[1] 黄强[1] 董军[1] 宋武超[1] 王爱东[1] 兰青[1] 

机构地区：[1]苏州大学附属第二医院神经外科暨脑肿瘤研究室,215004

出　　处：《中国现代神经疾病杂志》2009年第6期544-547,共4页Chinese Journal of Contemporary Neurology and Neurosurgery

基　　金：国家自然科学基金资助项目(项目编号:30672164);国家自然科学基金资助项目(项目编号:30772241);江苏省青年科技创新人才项目(项目编号:BK2007507)

摘　　要：目的探讨建立脑肿瘤裸小鼠原位移植模型的方法。方法于脑立体定向仪辅助定位下,分别将经体外培养的脑肿瘤干细胞球(SU-1和SU-2)和人多形性胶质母细胞瘤细胞系单细胞悬液缓慢注射至裸小鼠右侧尾状核;徒手将SHG44细胞皮下移植瘤组织块、人多形性胶质母细胞瘤体外细胞系移植成功获得的颅内肿瘤组织块和肺癌脑转移瘤组织块接种于裸小鼠右侧尾状核,HE染色检测不同来源肿瘤细胞致瘤率。结果细胞悬液注射法致瘤率为100%(45/45)。肿瘤组织块直接移植法致瘤率分别为:SHG44细胞皮下移植瘤为93.33%(14/15),人多形性胶质母细胞瘤体外细胞系移植瘤为98.46%(128/130),肺癌脑转移瘤为72.31%(47/65);裸小鼠死亡率为1.41%(3/213)。形成的可移植性肿瘤分为侵袭性明显和不明显两种类型,与亲本肿瘤特征一致。结论两种移植方法均可成功制备包括脑肿瘤干细胞在内的人胶质瘤裸小鼠原位移植模型,反映了亲本肿瘤最基本的致瘤性和侵袭性特征。细胞悬液注射法操作时间较长,但为体外培养的单个细胞和细胞球移植所必须;肿瘤组织块直接移植法可用于临床肿瘤、可移植性肿瘤的直接接种,操作时间短、效率高,适用于批量实验。Objective To study and compare the methods of establishment of brain tumor orthotopic transplantation model in nude mice. Methods Two methods were developed to establish orthotopie transplantation model of nude mice. One of the methods was direct injection of brain tumor stem cells （BTSCs） suspension or glioblastoma （GBM） cells suspension into the caudate nuclei of nude mice under the guidance of stereotaxic apparatus. Another was tumor tissue （SHG44 subcutaneous xenografts, GBM intracranial xenografts and brain metastasis of human lung adenocarcinoma） transplantation via trocar and direct injection into the caudate nuclei of nude mice. Results Success rate of establishment of brain tumor orthotopic transplantation model was 100% with the first method. The success rate of the second method by transplanted with SHG44 subcutaneous xenografts was 93.33% （14/15）, GBM cells intracranial xenografts was 98.46% （128/130）, and brain metastasis of human lung adenocarcinoma was 72.31% （47/65）. The overall mortality rate of the second method was 1.41% （3/213）. According to the origin tumor which was transplanted, the tumor developed in brain of the mice was classified into invasive and non-invasive type. Conclusion Both methods can be used to develop brain orthotopic transplantation model, and the tumorigenicity and invasive features of the origin tumor were faithfully maintained in the models with both methods. Though the operative time is much longer, the first method is necessarily for cultivate single cell suspension and BTSCs spheres in vitro. The second method can be used when direct tumor tissue transplantation is performed, obviously it also bear advantage of time saving and easy to manipulate, and can be performed in large scale.

关 键 词：脑肿瘤 神经胶质瘤 干细胞 异种移植模型抗肿瘤试验 小鼠 裸 疾病模型 动物 

分 类 号：R739.4[医药卫生—肿瘤]