Direct comparison of five different 3D extracellular matrix model systems for characterization of cancer cell migration

Abstract

Background: Three-dimensional (3D) in vitro model systems can bridge the gap between regular two-dimensional cell culture and whole-animal studies. Analyses of cancer cell migration and invasion increasingly use differing 3D systems, which may produce conflicting findings. Aims: We directly compared different 3D extracellular matrix systems for studying cancer cell migration/invasion by analyzing cell morphologies and quantifying aspects of cell migration including speed and directional persistence using automated computer-based cell tracking. Methods and results: We performed direct comparisons of five different 3D extracellular matrix cell culture systems using both HT1080 fibrosarcoma and MDA-MB-231 breast carcinoma cell lines. The reconstituted 3D systems included two types of collagen hydrogel and tissue matrix gel (TMG) vs cell-derived matrices extracted from cultured primary human or cancer-associated fibroblasts. The fibrillar matrix architecture of these systems differed. 3D rat tail collagen and TMG matrices had short, randomly oriented collagen fibrils; bovine collagen had long, larger fibril bundles; and the cellderived matrices were strongly oriented. HT1080 cells displayed rounded morphologies in all three reconstituted 3D matrices but became spindle shaped in the two cell-derived matrices. MDA-MB-231 cell morphologies were elongated in all matrices. Quantitative measures of cell migration parameters differed markedly between the different types of 3D matrix. Comparing the reconstituted matrices, cells migrated the most rapidly and furthest in TMG. Comparing TMG with cell-derived matrices, cells migrated more efficiently in the cell-derived matrices. The most notable differences were in directional persistence of migration, which was greatest in the two cell-derived matrices. Conclusion: The morphologies of matrix fibrils and cell shape, and particularly the efficiency and directionality of cell migration, differed substantially depending on the type of 3D matrix system. We suggest that it is important to employ the 3D model system that most closely resembles the matrix environment being studied for analyses of cancer cell migration and invasion.

Inhomogeneities in 3D Collagen Matrices Impact Matrix Mechanics and Cancer Cell Migration

“…Collagen fibers from tendon are described as crimped and more uniform as the even fibers of the skin and thus exhibit different mechanical properties (Meyer, 2019) matched to the tissues they are part of. One major difference in the properties of reconstituted collagen I fibrillary networks from tendon or skin occurs due to the different extraction methods that are used to obtain collagen monomers (Demou et al, 2005;Wolf et al, 2009;Shinsato et al, 2020). More cross-linked skin collagens were additionally pepsinized to cleave telopeptides.…”

Keloid disorder: Fibroblast differentiation and gene expression profile in fibrotic skin diseases

“…All published 3D in vitro and animal models duplicate some but not all features of keloid development, and consequently, none can be considered as the “gold standard” for all investigations. A recent report by Shinsato et al, in which five different 3D ECM model systems were used to characterize differences in cancer cell migration, concluded that the morphology of matrix fibrils and cell shape are dependent upon the type of 3D matrix system utilized 121 . These matrix differences translate into changes in the migration/invasion properties of the resident cells.…”

Organ-Specific, Fibroblast-Derived Matrix as a Tool for Studying Breast Cancer Metastasis

“…Increasingly, 'simpler' ECM components have also been the focus of cancer studies to include a relevant tumour microenvironment. However, they are often limited in their ability to recapitulate these cancer niche properties as products such as Matrigel and gelatin are only composed of a limited number of ECM components and lack resemblance to native tissue architecture and biophysical properties [51]. The presence of the ECM is also critical when studying cancer cell behaviour in vitro to aid in increasing the efficacy of drugs tested in vitro to patients, as current studies suggest fewer than 5% of all drugs tested make it to the clinic [52,53].…”