RESUMO
Microfluidic cell cultures enable investigation of complex physiological tissue properties and functionalities. For convenience, they are often implemented with immortalized cell lines, but primary cells more closely approximate the in vivo biology. Our aim was to develop a biomimetic microfluidic model of the human airway using all primary cells. The model is comprised of airway epithelial cells cultured at an air-liquid interface, lung fibroblasts and polarized microvascular endothelial cells, respectively positioned in three vertically stacked, individually accessible compartments separated by nanoporous membranes. We report device fabrication, a gravity fed microfluidic system, and culture medium able to support functional co-cultures of all three primary human cell types. As characterized by imaging and permeability measurements, airway epithelial cells in microfluidic devices displayed mucociliary differentiation and barrier function. Subjacent fibroblasts and microvascular endothelial cells were added under conditions enabling co-culture for at least 5 days. Microfluidic airway models based on primary human cells in a relevant biomimetic configuration will improve physiological relevance and will enable novel disease modeling and drug development studies.
