Three-dimensional tumor-stroma co-culture system development using self-assembling peptide scaffolds

Cancer research has traditionally relied on 2D cell culture, focusing mainly on cancer cells and its disrupted genetics. However, tumors have been accepted as complex tissues, and as such, they need signals from a 3D environment to form tissue structures in vitro. Moreover, cancer cells behavior can only be deciphered considering the contribution of the cells existing in the tumor stroma as well as its complex microenvironment. Since the tumor microenvironment plays an important role in cancer progression, it is widely accepted that culturing cells in 3D scaffolds, which mimic the native extracellular matrix, represents a more realistic scenario. In the present work we aim to develop an in vitro 3D co-culture system that would comprise both cancer and stromal cells. For that, HeLa cells were injected into a RAD16-I peptide scaffold containing fibroblasts, resulting in a 3D system were cancer cells were embedded within a stromal cells matrix. With this system, we were able to study cancer cells behavior in a 3D context in terms of survival, migration and proliferation. Moreover, we have demonstrated that the anti-cancer effect of different pharmaceutical drugs (Gemcitabine, 5-Fluorouracil and the ROCK inhibitor Y-27632) can be qualitatively and quantitatively evaluated on the 3D co-culture system developed.