Environmental and Chemical Engineering School, Dalian University, Dalian 116622, PR China; Microfluidic Research Institute, Dalian University, Dalian 116622, PR China.
Medical School, Dalian University, Dalian 116622, PR China; Microfluidic Research Institute, Dalian University, Dalian 116622, PR China.
Exp Parasitol. 2014 Feb;137:1-7. doi: 10.1016/j.exppara.2013.11.009. Epub 2013 Dec 6.
In vitro culturing of trophozoites was important for research of Giardia lamblia (G. lamblia), especially in discovery of anti-Giardia agents. The current culture methods mainly suffer from lab-intension or the obstacle in standardizing the gas condition. Thus, it could benefit from a more streamlined and integrated approach. Microfluidics offers a way to accomplish this goal. Here we presented an integrated microfluidic device for culturing and screening of G. lamblia. The device consisted of a polydimethylsiloxane (PDMS) microchip with an aerobic culture system. In the microchip, the functionality of integrated concentration gradient generator (CGG) with micro-scale cell culture enables dose-response experiment to be performed in a simple and reagent-saving way. The diffusion-based culture chambers allowed growing G. lamblia at the in vivo like environment. It notable that the highly air permeable material of parallel chambers maintain uniform anaerobic environment in different chambers easily. Using this device, G. lamblia were successfully cultured and stressed on-chip. In all cases, a dose-related inhibitory response was detected. The application of this device for these purposes represents the first step in developing a completely integrated microfluidic platform for high-throughput screening and might be expanded to other assays based on in vitro culture of G. lamblia with further tests.
体外滋养滋养体对于研究蓝氏贾第鞭毛虫(G. lamblia)非常重要,特别是在发现抗贾第鞭毛虫药物方面。目前的培养方法主要受到实验室强度或标准化气体条件的障碍的影响。因此,需要更精简和集成的方法。微流控技术为此提供了一种途径。在这里,我们提出了一种用于培养和筛选蓝氏贾第鞭毛虫的集成微流控装置。该装置由具有需氧培养系统的聚二甲基硅氧烷(PDMS)微芯片组成。在微芯片中,集成浓度梯度发生器(CGG)与微尺度细胞培养的功能使剂量反应实验能够以简单且节省试剂的方式进行。基于扩散的培养室允许蓝氏贾第鞭毛虫在类似于体内的环境中生长。值得注意的是,平行室的高透气性材料可轻松维持不同室中的均匀厌氧环境。使用该装置,成功地在芯片上培养和应激蓝氏贾第鞭毛虫。在所有情况下,都检测到与剂量相关的抑制反应。该设备在这些用途上的应用代表了开发用于高通量筛选的完全集成微流控平台的第一步,并且可以通过进一步的测试扩展到其他基于蓝氏贾第鞭毛虫体外培养的测定。