Towards an immunocompetent cervix on-chip infection model: endothelialization and optimization of macrophage culture.

The human cervix, representing the lower part of the female uterus, is highly susceptible to bacterial and viral infections leading to possible severe health outcomes. Only limited prevention and treatment options are currently available for patients, which is partially due to the lack of human-relevant preclinical models. Microfluidic cervix-on-chip platforms are promising tools to study cervical infections in a human-specific manner in order to test potential drug and vaccine candidates in the future. In this master thesis project, an existing cervix-on-chip platform was refined towards an endothelialized immunocompetent model allowing the investigation of infection with pathogens like Herpes simplex virus 2. For this purpose, we established and optimized isolation and culture conditions of primary human cervical endothelial cells. We further evaluated the functionality of cervical endothelial cells through tumor necrotic factor alpha stimulation. Endothelial cells activation revealed changes in the expression of specific junctions proteins and immunomodulatory adhesion molecules, indicating physiological responses of these cells to inflammatory stimuli present also in the context of infection. Subsequently, we assessed ideal on-chip culture conditions and established a cervical endothelial cell barrier on-chip. To a greater extent, this work lays the foundation for integrating tissue-resident macrophages into the cervical tissue model. We isolated and differentiated blood-derived monocytes into macrophages, thereafter we emulated on-chip culture conditions on-plate in order to examine macrophage viability. The obtained results could further be used to integrate both endothelial cells and tissue-resident macrophages on-chip. Moreover, the interplay between endothelial cells and immune cells could be investigated to study how the response to viral infections is modulated in the specific and complex context of the human uterine cervix. Overall this research represents an important contribution to gynecological infection models.