13. NF-KAPPAB SIGNALING IN A DYNAMIC MICROFLUIDIC ENVIRONMENT

Department: Bioengineering
Faculty Advisor(s): Jeff Hasty

Primary Student
Name: Martin Kolnik
Email: mkolnik@ucsd.edu
Phone: 858-534-5594
Grad Year: 2012

Abstract
Nuclear factor kappa B is a well-studied global regulator of gene expression that coordinates the cellular response to a variety of external stimuli such as stress and infection. NFkappaB is normally sequestered in the cytoplasm but it translocates into the nucleus upon stimulation and acts to regulate a variety of downstream genes before it is shuttled out of the nucleus back into the cytoplasm. Oscillation dynamics of NFkappaB shuttling have been implicated in the functional dynamics of subsequent gene expression but it remains to be determined to what extent dynamic stimulation of the system affects nuclear-cytoplasmic NFkappaB shuttling. To this end, we are developing a microfluidic cell culture device to stimulate mammalian cells with any desired time-varying waveform of biochemical inducer while maintaining the cells in a zero-shear environment. This is accomplished by isolating the cells away from the main flow channel of the device in square-shaped "side traps" which have only one side open to the main channel thereby creating an extremely high resistance to flow inside the cell traps. This ensures that diffusive transport rather than convective forces is the dominant means of nutrient and biochemical inducer exchange. A previously developed microfluidic mixer known as the "Dial-a-Wave" is then used to generate the desired waveform of the biochemical inducer TNFalpha which is used to stimulate the NFkappaB cellular response.

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