Investigation on the Composition of Agarose–Collagen I Blended Hydrogels as Matrices for the Growth of Spheroids from Breast Cancer Cell Lines

Abstract

Three-dimensional (3D) cell culture systems mimic the structural complexity of the tissue microenvironment and are gaining increasing importance as they resemble the extracellular matrix (ECM)–cell and cell–cell physical interactions occurring in vivo. Several scaffold-based culture systems have been already proposed as valuable tools for large-scale production of spheroids, but they often suffer of poor reproducibility or high costs of production. In this work, we present a reliable 3D culture system based on collagen I-blended agarose hydrogels and show how the variation in the agarose percentage affects the physical and mechanical properties of the resulting hydrogel. The influence of the different physical and mechanical properties of the blended hydrogels on the growth, size, morphology, and cell motility of the spheroids obtained by culturing three different breast cancer cell lines (MCF-7, MDA-MB-361, and MDA-MB-231) was also evaluated. As proof of concept, the cisplatin penetration and its cytotoxic effect on the tumor spheroids as function of the hydrogel stiffness were also investigated. Noteworthily, the possibility to recover the spheroids from the hydrogels for further processing and other biological studies has been considered. This feature, in addition to the ease of preparation, the lack of cross-linking chemistry and the high reproducibility, makes this hydrogel a reliable biomimetic matrix for the growth of 3D cell structures.

Targeting Cancer Cell Tight Junctions Enhances PLGA-Based Photothermal Sensitizers’ Performance In Vitro and In Vivo

“…Since the efficacy of both low molecular weight compounds and supramolecular structures differs significantly within in vitro and in vivo models, 3D spheroids are often used as an intermediate step as a more relevant tool for studying drug efficacy [21][22][23][24][25]. In particular, in spheroids, cells form tight intercellular contacts, which are often the main reason for the poor penetration of therapeutic substances into the tumor.…”

3D hydrogel-based microcapsules as an in vitro model to study tumorigenicity, cell migration and drug resistance

“…When analyzing the progression of breast cancer pathologies in vitro, it is relevant to note however, the wide number of studies still carried out on flat surfaces, or with cells entrapped in very soft scaffolds [7][8][9][10][11][12]. These strategies, however, are mismatching the morphology of solid tumors, or the elastic properties already reported for malignant tissues (between 15 to 80 kPa in the case of breast cancer, depending on the progression of this pathology) [13].…”

A thermo-sensitive chitosan/pectin hydrogel for long-term tumor spheroid culture

“…Both biological and synthetic biomaterials have been proposed to support the formation and growth of several types of cells and spheroids, as extracellular matrix (ECM)-like analogues, driving a variety of cellular processes (Kamatar et al, 2020;Quarta et al, 2021). Chemically defined synthetic polymers have been largely used to develop hydrogels systems for 3D culture (Aisenbrey & Murphy, 2020).…”