Understanding the Impact of 2D and 3D Fibroblast Cultures on In Vitro Breast Cancer Models

Kyung Eun Sung,Carolyn Pehlke,Xiaojing Su,Erwin Berthier,David J Beebe,Andreas Friedl,Edna Cukierman

doi:10.1371/journal.pone.0076373

Kyung Eun Sung, Carolyn Pehlke + Show 5 more

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https://doi.org/10.1371/journal.pone.0076373

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Abstract

The utilization of 3D, physiologically relevant in vitro cancer models to investigate complex interactions between tumor and stroma has been increasing. Prior work has generally focused on the cancer cells and, the role of fibroblast culture conditions on tumor-stromal cell interactions is still largely unknown. Here, we focus on the stroma by comparing functional behaviors of human mammary fibroblasts (HMFs) cultured in 2D and 3D and their effects on the invasive progression of breast cancer cells (MCF10DCIS.com). We identified increased levels of several paracrine factors from HMFs cultured in 3D conditions that drive the invasive transition. Using a microscale co-culture model with improved compartmentalization and sensitivity, we demonstrated that HMFs cultured in 3D intensify the promotion of the invasive progression through the HGF/c-Met interaction. This study highlights the importance of the 3D stromal microenvironment in the development of multiple cell type in vitro cancer models.

Highlights

Cancer cells cultured in an extra cellular matrix (ECM) (often called three-dimensional (3D) culture) show differences in functional behaviors such as differentiation, proliferation, and gene expression [1,2,3], when compared to cells cultured on a flat surface (two-dimensional (2D))
We focus on examining the paracrine interaction between cancer cells and stromal fibroblasts during the breast cancer progression from ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) in the context of matrix effects on the stromal cells and their subsequent regulation of cancer progression
We examined the influence of 2D and 3D culture of human mammary fibroblasts (HMFs) on the invasive transition of breast cancer cells (MCF10-DCIS.com (MCFDCIS) cells), known as the DCIS to IDC transition

Summary

Introduction

Cancer cells cultured in an extra cellular matrix (ECM) (often called three-dimensional (3D) culture) show differences in functional behaviors such as differentiation, proliferation, and gene expression [1,2,3], when compared to cells cultured on a flat surface (two-dimensional (2D)). The growing consensus is that 3D models recreate key aspects of the microenvironment more faithfully and, in some cases, provide more comprehensive and relevant biological information that is impossible or difficult to obtain from 2D models [4,5,6] This realization has prompted increased use and exploitation of 3D culture for in vitro cancer models [3,7,8,9]. One hypothesis attributes the changes observed in 3D culture to the enhanced interactions between cells and the surrounding ECM This hypothesis is supported by reports of a growing number of different signaling mechanisms in 3D microenvironments compared to 2D microenvironments over the last decade [7,9,10,11,12]. We focus on examining the paracrine interaction between cancer cells and stromal fibroblasts during the breast cancer progression from DCIS to IDC in the context of matrix effects on the stromal cells and their subsequent regulation of cancer progression

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