Development of a 3D Co-Culture System as a Cancer Model Using a Self-Assembling Peptide Scaffold.

Nausika Betriu,Carlos Semino

doi:10.3390/gels4030065

Abstract

Cancer research has traditionally relied on two-dimensional (2D) cell culture, focusing mainly on cancer cells and their abnormal genetics. However, over the past decade, tumors have been accepted as complex tissues rather than a homogenous mass of proliferating cells. Consequently, 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 a critical role in tumorigenesis, it is widely accepted that culturing cells in three-dimensional (3D) scaffolds, which mimic the extracellular matrix, represents a more realistic scenario. In the present work, an in vitro 3D co-culture system based on the self-assembling peptide scaffold RAD16-I (SAPS RAD16-I) was developed as a cancer model. For that, PANC-1 cells were injected into a RAD16-I peptide scaffold containing fibroblasts, resulting in a 3D system where cancer cells were localized in a defined area within a stromal cells matrix. With this system, we were able to study the effect of three well-known pharmaceutical drugs (Gemcitabine, 5-Fluorouracil (5-FU), and 4-Methylumbelliferone (4-MU)) in a 3D context in terms of cell proliferation and survival. Moreover, we have demonstrated that the anti-cancer effect of the tested compounds can be qualitatively and quantitatively evaluated on the developed 3D co-culture system. Experimental results showed that Gemcitabine and 5-FU prevented PANC-1 cell proliferation but had a high cytotoxic effect on fibroblasts as well. 4-MU had a subtle effect on PANC-1 cells but caused high cell death on fibroblasts.

Highlights

An in vitro 3D co-culture system based on the self-assembling peptide scaffold RAD16-I
Peptide scaffold containing fibroblasts, resulting in a 3D system where cancer cells were localized in a defined area within a stromal cells matrix
Prevented PANC-1 cell proliferation but had a high cytotoxic effect on fibroblasts as well. 4-MU had a subtle effect on PANC-1 cells but caused high cell death on fibroblasts

Summary

Introduction

Cancer research has relied both on two-dimensional (2D) cell culture and on animal models. Animal models are a valuable tool in cancer research. They do not represent the behavior of naturally occurring cancers in humans and are expensive, time-consuming, and not feasible for high-throughput screening (HTS) [1,2,3]. 2D in vitro cancer models are under highly controlled conditions and are highly reproducible, which make them very attractive to be used routinely for many research groups and the pharmaceutical industry. It is widely accepted that 2D models do not represent the in vivo scenario, since cells are grown in a solid and flat support. When culturing cells in 2D, the extracellular matrix (ECM)’s composition and configuration are strongly modified, and cells do not receive the proper signals that provide a normal ECM configuration [4]

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Results