Fundamental studies of liver cell culture microtechnology and techniques for the purpose of designing a liver model

Abstract

An authentic human liver model has the potential to be a revolutionary development in the field of biotechnology, and the progress of microtechnology has made this idea a reality. This technology enables the construction of micro-environments that can closely mimic the architecture and environment of human liver tissue in vivo, which is the key to building a physiologically relevant model. To this end, novel platforms have been produced with the goal of eventually building an authentic human liver model. A simple, single channel has been used for preliminary work with primary rat hepatocyte culture. Despite some successes, this system has shown several flaws, which include leakage and inconsistent viability. In order to rectify the leakage issue, a novel clamping system was devised, and cell culture tests have illustrated its capability and reliability in preventing leakage. Additionally, an initially open channel system was devised as an attempt to solve viability issues. We have also developed a novel dual channel system for co-culture of both types of liver cells, and initial tests are promising. This platform is capable of supporting this co-culture, although not consistently. Primary hepatocyte cultures in this system also led to an accidental discovery about the improved viability and cell contacts when cells are in tight confinement. Another important aspect in developing a liver model is obtaining physiologically relevant oxygen levels. To this end, oxygen diffusion simulations were constructed in COMSOL to determine the potential levels in the novel dual channel system. These results are promising and need to be validated through an oxygen fluorescence experiment. A perfusion system has been constructed for this experiment, and initial tests have been run to determine the proper amount of dye for an adequate fluorescent signal.M.S., Mechanical Engineering -- Drexel University, 201

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