Development of a microfluidic chip for perfusion three-dimensional cell culture and real-time dynamic cell growth analysis

• Main Authors: Che Wei Hsu, 許哲瑋
• Other Authors: K. F. Lei
• Format: Others
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/23123445306536155343

碩士 === 長庚大學 === 醫療機電工程研究所 === 101 === A perfusion three dimensional (3D) cell culture microfluidic chip has been developed for the real-time and non-invasive impedimetric monitoring of cell dynamics. The chip can continuously estimate the cell proliferation or chemosensitivity under different culture conditions. It is capable of onsite detecting the cell number or viability in the 3D cell culture construct without sacrificing the cultured cells. The chip consists of a substrate, a cover layer and a fluidic layer. The chip substrate is a glass substrate and a pair of 3D vertical electrodes is fabricated on its surface. The cover layer and fluidic layer are made of poly¬dimethylsiloxane (PDMS) material and formed by soft lithography. A culture chamber is defined by the openings of the cover layer and the fluidic layer provides fluidic connection for medium perfusion purpose. By bonding these layers, the microfluidic chip can be fabricated. For the fabrication of the 3D vertical electrodes, planar electrodes were first fabricated on the substrate by standard micro-fabrication techniques. Then, the cover layer was bonded to the substrate with the alignment of the culture chamber and the planar electrodes. By copper electroplating process, a pair of 3D vertical electrodes were grown from the planar electrodes and located at the opposite sidewalls of the culture chamber. In this study, human oral cancer cell-line (OEC-M1) were used and encapsulated in 3D agarose scaffold and cultured in the microfluidic chip. On-site impedance measurement was performed to estimate the cellular response, i.e., cell number and viability, in the 3D culture construct. Cell number in the 3D construct was shown to be proportional to the impedance magnitude of the entire construct. Therefore, perfusion 3D cell culture was performed for up to 5 days and cell proliferation can be monitored by the impedimetric analysis. Furthermore, real-time monitoring of cell viability under the perfusion of anti-cancer drug in different concentrations was conducted and the impedance magnitude was directly correlated with the cell viability. From the confirmation of the endpoint cell viability assays, a concentration-dependent effect was shown; however, the response of cell viability during the drug treatment was able to be traced by the impedance measurement. This work showed cell proliferation and chemosensitivity in 3D cell culture format can be monitored by impedance measurement. This microfluidic chip has a high potential to develop a useful analytical platform for cancer research.