Modeling the Role of Cancer-Associated Fibroblasts in Tumor Cell Invasion

SIMPLE SUMMARY: Accurate in vitro modeling of diseases is essential to making breakthrough and clinically relevant discoveries. Assays to examine the process of invasion—a classical hallmark of cancer—have evolved over the years to overcome shortfalls in their design and accommodate new knowledge in the field, such as the role of the tumor microenvironment (TME) in propagating this process. The goals of this review are two-fold: To walk through the tried-and-true plus novel and new invasion assays currently used in cancer research with a focus on those incorporating cancer-associated fibroblasts (CAFs), and to be a resource for researchers to find the correct invasion assays that suit their own unique needs and biological questions. ABSTRACT: The major cause of cancer-related deaths can be attributed to the metastatic spread of tumor cells—a dynamic and complex multi-step process beginning with tumor cells acquiring an invasive phenotype to allow them to travel through the blood and lymphatic vessels to ultimately seed at a secondary site. Over the years, various in vitro models have been used to characterize specific steps in the cascade to collectively begin providing a clearer picture of the puzzle of metastasis. With the discovery of the TME’s supporting role in activating tumor cell invasion and metastasis, these models have evolved in parallel to accommodate features of the TME and to observe its interactions with tumor cells. In particular, CAFs that reside in reactive tumor stroma have been shown to play a substantial pro-invasive role through their matrix-modifying functions; accordingly, this warranted further investigation with the development and use of invasion assays that could include these stromal cells. This review explores the growing toolbox of assays used to study tumor cell invasion, from the simple beginnings of a tumor cell and extracellular matrix set-up to the advent of models that aim to more closely recapitulate the interplay between tumor cells, CAFs and the extracellular matrix. These models will prove to be invaluable tools to help tease out the intricacies of tumor cell invasion.