Title:

Mechanically Tunable Hydrogel-Based Clearance Assay for Ovarian Cancer Modeling

Late-stage cancer often is accompanied by metastasis, which is the leading cause of cancer-related deaths and treatment failure. Roughly 75% of ovarian cancer cases are diagnosed after metastasis has occurred. Ovarian cancer metastasis involves cancer cells migrating from the original tumor site, employing a mechanism that clears peritoneal mesothelial cells to invade new tissue. Many in vitro cancer models lack biologically relevant environments that reflect in vivo conditions. This research uses a Dextran Methacrylate (DexMA) hydrogel model to replicate in vivo tissue stiffnesses for use as a mesothelial clearance assay. DexMA and adhesion peptides were cured under UV light to create a range of stiffnesses. Nanoindentation was conducted to determine optimal UV cure times and DexMA concentrations. A concentration of 30mg/ml DexMA and UV cure times of 10, 20, 30, and 60s were found to produce an ideal range of stiffnesses. Stained LP9 mesothelial cells were cultured to form a monolayer on the surface of the gels, then OVCAR8 spheroids were placed on top. These samples were imaged at 0 and 24 hours to observe LP9 clearance by OVCAR8 spheroids. Using the hydrogel model, successful clearance was observed. This research provides a tunable hydrogel model for clearance assays.

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