研发了一种聚二甲基硅氧烷-纸复合型微流控芯片用于肝癌细胞三维培养。芯片使用明胶处理硝酸纤维素薄膜作为细胞培养基底,以水凝胶网格作为三维培养支撑。结合微通道主动灌流与水凝胶中的被动扩散,模拟体内的流体运输形式实现细胞与外界物质交换。实验结果显示,芯片上的液滴生成以及细胞定位种植简便可靠。连续监测显示肝癌HepG2细胞在水凝胶微球中增殖形成类似组织的三维结构。细胞增殖动力学分析以及生化检测结果显示了芯片三维培养与二维培养的差别。这种芯片三维细胞培养方法操作简便可靠,仿真度高,适合于肿瘤细胞研究。
We developed a Polydimethylsiloxane (PDMS)-paper hybrid microfluidic chip for three- dimensional (3D) culture of HepG2 cell, in which gelatin-treated nitrocellulose membranes were used as base of cell culture, and hydrogel grids were generated to serve as skeleton for 3D cultures. The fluid transportation combined active perfusion along the micochannel and passive diffusion through the extracellular matrix, which closely mimic the exchange of extraeellular fluid and mieroenvironmental substance in-vivo. Experimental results demonstrated that droplet generation, cell seeding and cell localization on the paper-embedded microchip can be achieved easily and reliably. Under continuous perfusion and monitoring, the liver cancer cells cultured in 3D matrix proliferated and spontaneously aggregated to form tissue-like 3D structures. Studies have shown that the cells grown in 3D differ considerably compared to 2D, proved by growth dynamics and biochemical tests. The microchip-based 3D cell culture features simplicity and precise emulation of the microenvironment in-vivo, and is thus well suited to various kinds of tumor cell research.