In vitro models for airway epithelial cell culture

Abstract

This work is about the development of a physiologically relevant model of the human airway. Various factors such as the cell model, physiochemical factors such as the cell substrate properties including its stiffness, shear stress, stretch, the air-liquid interface and the biochemical factors in the medium influence the biology of the cells. The aim of this work is to closely approximate conditions in an in vivo situation by engineering the above conditions in to the in vitro platform. An assay to introduce the cell substrate properties was developed in a glass bottomed petri dish type culture as well as a microfluidic device culture. The influence of the cell substrate on airway epithelial cell monolayer formation was investigated in detail by changing the stiffness of the substrate independently by changing the gel concentration, the gel formation pH and the height of the gel from a hard substrate. Further, we found that biochemical growth factors have a huge role in cell monolayer formation. A real-time measurement of monolayer integrity using electrical resistance measurements was developed. A shear stress application platform was developed and a stretch application platform was designed. The applications of such a platform with the inclusion of various physiologically relevant factors include the study of physiologic evolution of microbes such as the influenza virus.