"Exploring Mechanically-Induced Signaling in Kidney Epithelial Tissue: A Dual-Sided Perfused Flow Chamber Approach for Extended Cell Culture and Cellular Response Characterization"

Our lab uses cultured kidney epithelial tissue to better understand mechanically-induced signalling in ciliated epithelial cells. In the fields of cell & tissue physiology, epithelial and endothelial cells can be studied using a perfused flow chamber (‘FC’) which 1) allows researchers to investigate the direct effects of bulk shear stress on intact tissue and 2) mimics a physiologically relevant environment for cultured tissue. However, cell/tissue culture within the confined volume of a fluid chamber introduces numerous complications compared to ‘classical’ cell/tissue culture in petri dishes. For example, there is an absence of standard protocols to help grow cells, and the small volumes of media requires more attention be paid to the buildup of excreted metabolic byproducts. In addition, there are complex operational control and FC design requirements. We are designing a dual sided FC where cells are plated and grow on a semi-permeable membrane, allowing epithelial tissue to perform its function as a selective transporter of material into and out of the organ(ism). Because the tissue culture is exposed to medial on both sides (apical and basolateral), we have the option to expose the tissue to 2 different kinds of media (for example, a blood side and an ultrafiltrate side). In addition, as it is dual sided we have independent control over the fluid flow on apical and basolateral sides, allowing us to record cellular responses to a range of different fluid flow patterns.I will present the work I have done to date, showing how we can now culture cells within a flow chamber for many days, and also demonstrate some of the experimental techniques we will use to characterize cell responses.