Integrated Microfluidic Devices for Cell Culture and Assay

Citation

Liu, Mike Chia-Chung (2010) Integrated Microfluidic Devices for Cell Culture and Assay. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/D43B-D825. https://resolver.caltech.edu/CaltechTHESIS:10162009-053129363

Abstract

This thesis presents the development of three-dimensional (3-D) microfluidic devices for cellular studies, with focus on applications for high-throughput cell culture and cell-based assay. Microfluidic devices provide potential inexpensive platforms for high-throughput screening with the advantages of precise liquid handling, ability to control cell culture microenvironment, and reduced reagents and cells.

Because a mixture of drugs or chemical compounds can often treat diseases more effectively or act synergistically in certain cellular pathways, a device capable of screening the combinatorial effects of multiple compound exposures on cells is highly desirable. To this end, a novel method to monolithically fabricate 3-D microfluidic networks was developed, and based on this fabrication technology, the first cell culture device with an integrated combinatorial mixer was constructed. The proof-of-concept chip having a three-input combinatorial mixer and eight individually isolated micro culture chambers was fabricated on silicon utilizing the surface micromachining of Parylene C (poly(chloro-p-xylylene)). Unlike other 3-D microfluidic fabrications, multilayer bonding process was favorably obviated. By incorporating several microfluidic overpass structures to allow one microfluidic channel to cross over other microfluidic channels, the combinatorial mixer generated all the combinations of the input fluidic streams. Cell culturing on-chip was successful, and the ability to simultaneously treat arrays of cells with different combinations of compounds was demonstrated.

To facilitate cell-based assay, another combinatorial cell array device was fabricated on glass with incorporated membrane. Characterization of the combined compound concentration profile at each chamber with a fluorescence method was developed. We demonstrated functionality of the quantitative cell-based assay by screening three different compounds’ ability to reduce cytotoxicity of hydrogen peroxide on neuron cells and also assaying combinatorial exposures of three chemotherapeutic agents on breast cancer cells. The 3-D microfluidic fabrication process was extended to construct multilayer microfluidic device with integrated membrane. Applications of microfluidic devices for marine microbiology were demonstrated. Based on the capabilities demonstrated in this work, devices with high-density cell array and integrated high-input combinatorial mixer can be constructed. At the same time, the technology has general applicability for building complex 3-D microfluidic devices, which can broaden the applications for current lab-on-a-chip systems.

Thesis Files