Introduction/Background The cost of drug discovery is steadily increasing owing to the limited predictability of two-dimensional (2D) cell culture and animal models. The objective of this study was to develop a microfluidic ‘uterine cervix-on-a-chip’, a dynamic 3D platform that let the cultured cells mimic the native human uterine cervix histomorphology in vitro aiming to study the transformation zone of cervix during Human Papilloma virus (HPV) infection and cervical cancer development.
Methodology A microfluidic device prepared by demolding cured polidimetilsiloxano (PDMS). On the chip, we carried out cell culture and co-culture of ectocervical epithelial cells (Ect1/E6E7) and endocervical epithelial cells (End1/E6E7). Endocervix epithelial cells have been marked by AAV-GFP control viruses to provide a convenient way to measure transduction efficiency via fluorescence and to differentiate them from ectocervix epithelial cells. Live/dead assay, prestoBlue cell viability, 2D migration of cells (scratch test) and 3D migration of cells by the use of 3D printers were performed to check the functionality of the chip.
Results As a preliminary result, we have made a non-toxic, functional, in vitro model for human uterine cervix that allows both types of epithelial cells to grow on both sides of the chip to reach each other in order to make the squamo-columnar junction. This multilayer-cell junction can mimic the transformation zone of the cervix. The next phase of the project is to mimic the infection via the upper chamber and to test the new treatments on the infected cells in order to evaluate changes in cytokines and immune response cells, as well as HPV mechanism of action.
Conclusion Uterine cervix-on-a-chip may provide a powerful alternative in vitro model for studies on cervix physiology, real-time and high-resolution imaging, and analysis of biological responses in the cervix, as well as drug development.
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