Creation of a urothelial permeability culture chamber

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Project Overview

In health, the bladder epithelium (urothelium) forms a complex barrier that restricts the translocation of ions, solutes, and bacteria into the body despite large changes in the volume, tonicity and composition of urine. During urinary tract infections E. coli damages this urothelial barrier by altering tight junction integrity between the epithelial cells thereby increasing urothelial permeability and clinical morbidity. This barrier function loss is manifested as changes in measured voltage and current across the epithelium resulting in a decreased calculated transepithelial electrical resistance (TEER). Currently short-term (5-10h) mechanistic studies use Ussing chambers to measure changes in urothelial biopsy TEER while long-term (days) studies rely on cell culture models. Neither is ideal as the short-term studies do not provide adequate time for tight junction damage and repair while long-term cell culture studies are unable to reliably mimic urothelial architecture. Hence our laboratory has been attempting to adapt an organoid culture model (ref 1) for long-term TEER measurements. Unfortunately, urothelial explants contract unpredictably over time and when using Corning Transwells the scaffolding becomes exposed making tissue TEER measurements unreliable.
We are proposing to develop a multichamber system capable of fixing the urothelium at a constant diameter within a Transwell-like cell culture system. The system would need to be reuseable, able to be sterilized, able to maintain a healthy urothelium at a constant diameter, and provide the ability for accurate TEER measurements for 5d.

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Team members from left to right: Kate Griffin, Jahnavi Puranik, Haleigh Simon, Alyssa Walker, Jiayi Lin

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