Researcher(s)
- Emily Buckley, Biological Sciences, University of Delaware
Faculty Mentor(s)
- Filipa Ribeiro Soeiro de Carvalho, Biomedical Engineering, University of Delaware
Abstract
Validating an In Vitro Controlled Membrane Buckling Platform to Study Gut Physiology
By Emily Buckley, Filipa Ribeiro, Katherine M. Nelson, Jason P. Gleghorn
The small intestine is lined with villi, which are fingerlike projections that dramatically increase surface area to enhance nutrient absorption and support rapid epithelial renewal. The structure of the small intestine plays a significant role in how epithelial cells grow, organize, and migrate; however, replicating this structure is challenging in in vitro models. As a result, many experiments fail to recreate the physiology of the intestinal environment, which can affect cell behavior and function. Therefore, we developed an in vitro cell seeding platform to create controlled buckling of a polycarbonate membrane that better mimics the topography of intestinal villi. An insert consisting of a silicone channel cut to a desired shape was placed on the platform and stretched by 20%. After attaching a polycarbonate membrane to the silicone, we removed the insert from the platform, creating controlled buckles in the channel. We then sealed the channel with agarose gel to create a confined channel to seed epithelial cells. To confirm the integrity of the seal and the insert structure, a food dye solution was pipetted into the channel then visually inspected for leakage. Next, madin-darby canine kidney (MDCK) cells were seeded onto the insert at 100,000 cells per 30µL. Then, the cells were tested for their ability to adhere to the buckled membrane, form a complete monolayer barrier, and migrate to the peaks and valleys. The goal was to determine whether the platform could support cells through the sealing and buckling steps, allow them to adhere properly, and provide a suitable environment to observe directional migration in future experiments. The cells could successfully adhere to the curved membrane, remained viable, and the insert structure was maintained throughout the process. These outcomes suggest that the platform is compatible with cell culture workflows and has potential for studying how topography affects epithelial behavior. Future steps include seeding with Caco-2 cells, collecting imaging data, and comparing cell growth, organization, and migration across curved versus flat regions of the insert.