Development of paper-based 3D cell culture devices and real-time biosensing applications

Abstract

Due to its micro fibrous pores, paper has gained traction in the use as a scaffold for 3D cell culture. To study melanoma mechanism, screening the amount of melanin production and invasion study are major keys for drug development. In this work, we developed paper-based devices for culturing melanoma cells. Through the development process, we obtained the optimal parameters, which were applied for 3 different projects; 1) Anti-melanogenic effect screening, 2) Impedance-based E-screen assay, and 3) A membrane insert for real-time invasion assay. As a result of melanin production from melanocytes, the cells can be easily observed after a few days of culturing due to their black color. Fortunately, this color change can easily be visualized when melanocyte is cultured in the paper due to the white color of the paper. The paper was then applied for anti-melanogenic screening of the natural compounds with short analysis time and easy steps. In order to develop the real-time invasion assay, we primarily study the impedance characteristics of cells. The obtained impedance parameters were applied for real-time estrogen screening as a parallel project. Based on the estrogen receptor on breast cancer cells, the binding of estrogen can increase cell proliferation, resulting in the increased impedance magnitude. The estrogenic effect of the test compounds, Bisphenol-A and Irgarol 1051, were then monitored in real time. After that, we applied the obtained impedance parameters for the development of real-time invasion assay device using paper as a membrane insert. The concept was based on the blockage of invasive cells within paper pores, resulting in the increased impedance magnitude. The device was applied for real-time invasion study of melanoma cells under the treatment of IGF-1. Melanoma invasion could be tracked within 7 h after IGF-1 treatment. Thus, the invasion of melanoma cells was less likely affected by cell proliferation.