Microfluidic devices for studying early response of cytokine signaling

This thesis presents the design, fabrication, and characterization of a microfluidic device integrated with cell culture, cell stimulation, and protein analysis as a single device towards an efficient and productive cell-based assay development. In particular, this thesis demonstrates the feasibility of culturing human cancer cells in microliter-volume reactors in batch and fed-batch operations, stimulating the cell under well-controlled and reproducible conditions at early stages, and detecting the protein signals with an immunocytochemical (In-Cell Western) assay. These microfluidic devices take advantage of microfabrication techniques to create an environment suitable for cell culture, biomechanical and biochemical stimulation of cells, and protein detection and analysis. The microfluidic approach greatly reduces the amounts of samples and reagents necessary for these procedures and the required process time compared with their macroscopic counterparts. Moreover, the technique integrates unit operations, such as cell culture, stimulation, and protein analysis, in a single microchip. The microfluidic technique presented in this thesis correlates the space in the microchannels with the biological process time (cell stimulation time). Thus, a single experiment in one microfluidic device is capable of generating a complete temporal cell response curve, which otherwise would have required multiple experiments and manual assays by standard microwells and pipetting techniques. The developed method also provides high time-resolution and reproducible data for studies of cell signaling events, especially at early stages. These cell signaling events are difficult to be investigated by conventional techniques. The thesis reports the development not only of a cell population analysis method, but also of a single-cell detection and analysis technique to explore cell-to-cell variations.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Chemical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2008

Author dc:creator

• Ye, Linlin, Ph. D. Massachusetts Institute of Technology

Advisor dc:contributor.advisor

• Klavs F. Jensen.

• Chemical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/43218

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/43218