摘要
目的建立乳腺癌细胞的微流控芯片模型,模拟人体内部微环境三维结构,对乳腺癌细胞进行初步药效筛选。方法利用3D打印技术,制作双层带细胞培养池的微流控芯片,进行乳腺癌细胞培养,通过基质胶构架模拟人体微环境,在芯片上进行紫杉醇抗乳腺癌细胞MCF7药效实验,通过AO/EB染色,在激光共聚焦镜下观察细胞死亡比率,并与传统二维药筛实验进行对比。结果在施以10μL·h^(-1)流速的情况下,6 d后,芯片里的细胞在基质胶支架里形成三维腔体结构。细胞在培养池里培养24 h后,以10μL·h^(-1)流速加含有0.02μmol·m L^(-1)的紫杉醇培养基,处理24 h后,通过激光共聚焦荧光观察到有明显的细胞死亡。结论本实验构建的微流控芯片模型可以实现乳腺癌细胞的三维培养,并进行初步的药物筛选,为进一步实现"人体芯片"奠定基础。
OBJECTIVE To develop a microfluidic mammalian cell culture device with human breast cancer cells co-cultured with extracellular matrix（ ECM）,reconstitute the 3-dimension human body microenvironment and preliminarily conduct drug screening on the breast cancer chip. METHODS 3D Printing technology was utilized to build a two-layer microchip with cell culture reservoirs.The breast cancer cells（ MCF7） were cultured in the matrix gel structure which mimiced the 3-dimension human body microenvironment. The cell toxicity of paclitaxel（ PTX） on MCF7 cells was preliminarily observed using this microfluidic chip. With the use of AO / EB immunofluorescent staining and laser confocal microscopy,the cell death ratio induced by PTX was determined and compared with that determined by 2-dimension drug screening methods. RESULTS MCF7 cells cultured on the chip successfully formed a 3D cavity structure in the matrix after 6-day dynamic incubation. The flow rate was 10 μL·h^-1. After 24 h dynamic culture,the culture medium was changed to culture solution containing 0. 02 μmol·m L^-1PTX,and the incubation was continued for 24 h. Obvious cell death was detected under laser confocal microscopy. CONCLUSION The microfluidic chip developed in this study can successfully culture breast cancer cells in 3-dimension structure and perform drug screening,which lays a foundation for actualization of＂human-ona-chip＂.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2015年第24期2124-2129,共6页
Chinese Pharmaceutical Journal
基金
十二五科技重大专项资助项目(2013ZX09103002-005)
江苏省科技支撑计划资助项目(BE2014654)
关键词
微流控芯片
3D打印技术
三维培养
药物筛选
microfluidic chip
3D printing technology
3D cell culture
drug screen
