Development of a Microfluidic Perfusion 3D Cell Culture and Measurement Biochip for Continuous Monitoring 3D Cellular Dynamic Response under Varied Culture Conditions

Measurement of cell number and viability during cell culture is an important process to explore the cell proliferation under a specific culture condition. Currently, it is generally realized by directly counting cells microscopically, detecting the turbidity of a cell suspension optically, or indirectly quantifying cellular components during the 2D cell culture. However, these techniques are normally labor-intensive and time-consuming, or needed to sacrifice the cultured cells. Also, 2D cell culture cannot well mimic the native cellular microenvironment since animal cells natively inhabit in 3D environments. Therefore, microfluidic perfusion 3D cell culture with the on-site 3D impedance measurement of cell characteristics is proposed to tackle these limitations. In this proposal, a microfluidic perfusion 3D cell culture & measurement biochip is proposed for investigating the 3D cellular dynamic response under varied culture conditions based on the impedance measurement. A culture cavity (4×4×1mm3) on the biochip is designed and its two opposite sidewalls are 3D copper electrodes while the other two opposite sidewalls are made of isolated PMMA. Cells are encapsulated in agarose gel and cultured in 3D environment in the culture cavity. A steady and homogenous 3D culture environment is constructed for faithfully exploring the physiological responses of cells to extracellular conditions. Microfluidic perfusion 3D breast cancer cell culture is performed on the proposed biochip and cell number and viability are measured by the on-site 3D electrodes simultaneously and instantly. The measurement provides an on-site, non-contact, non-label, and non-destructive analysis of cell characteristics during the 3D cell culture process. Moreover, evaluation of the 3D cellular dynamic response to the culture conditions can be continuously monitored. The duration of this project is planned for 3 years. In the first year, design and fabrication of the 3D cell culture & measurement biochip will be conducted. Development of the 3D electrodes using electroplating technique in the biochip will be investigated. Also, perfusion 3D cell culture will be investigated to demonstrate the feasibility for mimicking native cellular microenvironment. In the second year, 3D impedance measurement will be performed using the developed biochip. Measurement influenced by different conductivity medium and estimation of cell number in the 3D culture construct will be investigated. In the third year, measurement of cell viability in the 3D culture construct will be explored. The result will be evaluated by the conventional method. Moreover, continuous monitoring of 3D cellular dynamic response under varied culture conditions will be demonstrated.

Project ID:PB10202-1467
External Project ID:NSC101-2221-E182-003-MY3