| Summary: | 碩士 === 國立成功大學 === 工程科學系碩博士班 === 95 === This study reports a new perfusion-based, micro three-dimensional (3-D) cell culture platform for drug testing using enabling microfluidic technologies. The platform provides the features of homogenous and well-defined culture environments, efficient medium delivery and “smart cells/ agarose (scaffold) loading mechanism”, allowing more precise and high-throughput 3-D cell culture-based assay or testing to be realized. In this work, the perfusion-based, micro 3-D cell culture platform is designed and is fabricated based on SU-8 lithography and PDMS (poly-dimethylsiloxane) replication processes. The miniaturization of the culture scale in the culture platform contributes favorably to a low chemical gradient culture environment. Also, two types of pneumatic-based medium pumping mechanisms (the “spider-web” peristaltic pneumatic micropumps system and “modified serpentine-shape” pneumatic micropump system associated with an air tank) were integrated into the micro 3-D cell culture platform to provide efficient and economical culture medium delivery mechanism. Moreover, the “smart cell/agarose (scaffold) loading mechanism” was proposed, allowing the cells/3-D scaffold loading process in one step and avoiding too much laborious works and error caused by manual loading. The results show that in all of the 15 pneumatic micropumps studied, the medium delivery mechanism is able to provide a uniform flow output ranging from 3.6 to 439.0 µl/hr for the “spider-web pneumatic micropump system” and 5.5 to 131 µl/hr for the “modified serpentine-shape” pneumatic micropump system associated with an air tank, respectively, depending on the applied pulsation frequency of the micropumps. In addition, the cell/agarose (scaffold) loading mechanism is proven to be able to perform sample loading tasks precisely and accurately in all of the 15 microbioreactors investigated with adjustable loading volumes of 0.22, 0.18 and 0.14 µl at applied air pressures of 10, 12 and 15 psi, respectively, in the microvalves. Furthermore, anti-cancer drug testing is successfully demonstrated using the proposed culture platform and fluorescent microscopic observation. As a whole, because of miniaturization, not only does this perfusion 3-D cell culture platform provide a homogenous and steady cell culture environment, but it also reduces the need for human intervention. Moreover, due to the integrated pumping of the medium and the cell/agarose (scaffold) loading mechanisms, time efficient and economical research work can be achieved. These characteristics are found particularly useful for high-precision and high-throughput 3-D cell culture-based drug testing.
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