Development of a Paper-Based 3D Co-Culture Microfluidic System for Real-Time and Non-Invasive Impedimetric Monitoring of Cell-Cell Interaction under Various Cytokine Conditions

Epstein-Barr virus (EBV) is a human herpesvirus that is associated with several types of malignancy. EBV infects cells, stimulates their proliferation, induces the production of cytokines, and affects the aberrant proliferation of adjacent cells. Knowledge of this series of processes in cancers is essential for the development of effective therapeutic strategies. However, such investigation involves time consuming and labor intensive processes conventionally. That hampers the development of cancer research. Moreover, traditional two-dimensional (2D) cell culture cannot well mimic the native cellular microenvironment since animal cells natively inhabit in three-dimensional (3D) environments. Therefore, in order to tackle these limitations, a paper-based 3D co-culture microfluidic system is proposed to quantitatively monitor the cell-cell interaction under various cytokine conditions in a real-time and non-invasive manner. In this project, epithelial cells are firstly microinjected with pSG5-based expression vectors containing Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) (pSG5-LMP1). The transfected cells (LT-BM-1) and un-transfected cells (BM-1) are respectively encapsulated in hydrogel and the cell/hydrogel suspensions are respectively loaded to the co-culture system. The system mainly consists of a paper-based substrate with 2 adjacent culture regions and impedimetric measurement electrodes located at the bottom of the substrate. Cells are encapsulated in hydrogel and cultured in such paper-based 3D environment. Cytokines such as interleukin-6 (IL-6) are secreted from LT-BM-1 and affect the neighboring BM-1. Aberrant proliferation of BM-1 is induced. During the co-culture process, impedimetric monitoring for quantifying cell proliferation of both LT-BM-1 and BM-1 is simultaneously conducted and it is an on-site, real-time, non-contact, non-label, and non-destructive cellular analysis. In addition, neutralization and competitive assays are also performed to investigate the cell-cell interaction under various cytokine conditions. The proposed system provides a well-defined co-culture 3D environment and quantitative monitoring of cellular responses for the investigation of cell-cell interaction in order to develop effective therapeutic strategies. The duration of this project is planned for 3 years. In the first year, design and fabrication of the paper-based 3D co-culture microfluidic system will be conducted. Also, paper-based 3D cell culture will be investigated to demonstrate the feasibility for mimicking native cellular microenvironment. In the second year, on-chip monitoring of cellular responses of cell-cell interaction after virus infection will be investigated. In the third year, neutralization and competitive assays will be performed to study the cell-cell interaction under various cytokine conditions. These results are expected to understand more about how do cytokines affect the cell-cell interaction.

Project ID:PB10501-3721
External Project ID:MOST104-2221-E182-014-MY3