Department of Chemical Engineering, Texas A&M University, College Station, TX 77843-3122, USA.
Lab Chip. 2012 Apr 24;12(10):1813-22. doi: 10.1039/c2lc21202a. Epub 2012 Mar 28.
We describe the development of a fully automatic and programmable microfluidic cell culture array that integrates on-chip generation of drug concentrations and pair-wise combinations with parallel culture of cells for drug candidate screening applications. The device has 64 individually addressable cell culture chambers in which cells can be cultured and exposed either sequentially or simultaneously to 64 pair-wise concentration combinations of two drugs. For sequential exposure, a simple microfluidic diffusive mixer is used to generate different concentrations of drugs from two inputs. For generation of 64 pair-wise combinations from two drug inputs, a novel time dependent variable concentration scheme is used in conjunction with the simple diffusive mixer to generate the desired combinations without the need for complex multi-layer structures or continuous medium perfusion. The generation of drug combinations and exposure to specific cell culture chambers are controlled using a LabVIEW interface capable of automatically running a multi-day drug screening experiment. Our cell array does not require continuous perfusion for keeping cells exposed to concentration gradients, minimizing the amount of drug used per experiment, and cells cultured in the chamber are not exposed to significant shear stress continuously. The utility of this platform is demonstrated for inducing loss of viability of PC3 prostate cancer cells using combinations of either doxorubicin or mitoxantrone with TRAIL (TNF-alpha Related Apoptosis Inducing Ligand) either in a sequential or simultaneous format. Our results demonstrate that the device can capture the synergy between different sensitizer drugs and TRAIL and demonstrate the potential of the microfluidic cell array for screening and optimizing combinatorial drug treatments for cancer therapy.
我们描述了一种完全自动和可编程的微流控细胞培养阵列的开发，该阵列集成了芯片上药物浓度的产生和两种药物的成对组合以及平行培养细胞，用于药物候选物筛选应用。该设备具有 64 个可单独寻址的细胞培养室，细胞可以在其中进行培养，并依次或同时暴露于两种药物的 64 种成对浓度组合中。对于顺序暴露，使用简单的微流控扩散混合器从两个输入中产生不同浓度的药物。对于从两种药物输入生成 64 种成对组合，使用新的时间相关变量浓度方案与简单的扩散混合器结合使用，无需复杂的多层结构或连续介质灌注即可生成所需的组合。药物组合的生成和对特定细胞培养室的暴露使用能够自动运行多天药物筛选实验的 LabVIEW 接口进行控制。我们的细胞阵列不需要连续灌注来使细胞暴露于浓度梯度中，从而最小化每个实验使用的药物量，并且腔室内培养的细胞不会连续受到显著的剪切应力。该平台的实用性通过使用 doxorubicin 或 mitoxantrone 与 TRAIL（TNF-alpha Related Apoptosis Inducing Ligand）的组合，以顺序或同时的方式诱导 PC3 前列腺癌细胞丧失活力来证明。我们的结果表明，该设备可以捕捉不同敏化剂药物和 TRAIL 之间的协同作用，并展示微流控细胞阵列用于筛选和优化癌症治疗组合药物治疗的潜力。
