聚二甲基硅氧烷定制微培养装置及其在肿瘤细胞迁移浸润动态分析中的应用  

Development of customized PDMS apparatus and dynamic assays for cancer cell migratory invasion

作　　者：邢天禹[1,2] 牛静[1] 丁卫[1,2,3] 

机构地区：[1]首都医科大学基础医学院医学遗传学系,北京100069 [2]首都医科大学肿瘤研究所,北京100069 [3]北京市脑重大疾病研究院,北京100069

出　　处：《首都医科大学学报》2015年第2期270-275,共6页Journal of Capital Medical University

基　　金：国家自然科学基金(81372284;31201032)~~

摘　　要：目的利用新型有机硅材料聚二甲基硅氧烷(polydimethylsiloxane,PDMS)构建细胞组织微培养装置,建立肿瘤迁移浸润的动态分析方法,对迁移的细胞进行形态学和生物学检测。方法设计并用PDMS定制具有独立分割功能的细胞微培养装置。在人脑胶质瘤U87MG细胞中检测时间依赖的迁移行为和细胞迁移信号转化生长因子-β(transforming growth factor-beta,TGF-β)驱动的荧光素酶活性。检测肿瘤细胞在缺氧环境下迁移行为的改变,并与经典的Trans Well细胞迁移检测结果进行了对比。结果应用定制PDMS培养装置能够动态观测细胞迁移浸润,通过对迁移细胞群的分离回收的细胞和相关信号通路的快速定量分析能够更加全面准确地反映细胞迁移的动态表型改变和参与机制。结论应用PDMS定制细胞微培养装置进行细胞动态描述和分析,对研究肿瘤细胞迁移过程及其机制是一个可行且极具潜力的新方法。本微培养装置的特色在于在肿瘤细胞迁移过程中同时完成对形态学和分子生物学的分析并进行相关性研究。Objective To develop assays for the dynamic analyses for the migratory invasion of tumor through biocompatible polydimethylsiloxane（ PDMS） materials to produce tissue culture apparatus, thus to allow simultaneous morphological and biochemical measurements. Methods An adaptive PDMS mini-culture apparatus with a separate blade was designed and manufactured. U87MG glioblastoma cells were analyzed for time-dependent migration in correlation with the transforming growth factor-beta （ TGF-β） luciferase reporter activities. The cell behavior in response to hypoxia treatment was investigated and compared with the results from classic TransWell assays. Results The dynamic monitoring and determination of cell invasiveness and migration could be conveniently achieved with flexibility using the customized PDMS apparatus. By separation and collection of the subpopulations of high-mobility cells, the involved signal transduction pathways could be detected with ease and precision. Conclusion The dynamic characterization and analyses using PDMS-based customized cell culture devices can be an attractive approach to investigate the process and underlining mechanisms of cancer cell migration. The exemplary apparatus demonstrated in this study showed promising potentials for the coordinated investigation from both morphological and biochemical measurements during the tumor cell migration process.

关 键 词：聚二甲基硅氧烷 胶质瘤 迁移浸润 缺氧 转化生长因子Β 

分 类 号：Q-3[生物学] Q28