Brain Cancer Cell-Derived Matrices and Effects on Astrocyte Migration

Abstract

Cell-derived matrices are useful tools for studying the extracellular matrix (ECM) of different cell types and testing the effects on cell migration or wound repair. These matrices typically are generated using extended culture with ascorbic acid to boost ECM production. Applying this technique to cancer cell cultures could advance the study of cancer ECM and its effects on recruitment and training of the tumor microenvironment, but ascorbic acid is potently cytotoxic to cancer cells. Macromolecular crowding agents can also be added to increase matrix deposition based on the excluded volume principle. We report the use of macromolecular crowding (MMC) alone as an effective strategy to generate brain cancer cell-derived matrices for downstream analyses and cell migration studies. We cultured the mouse glioblastoma cell line GL261 for 1 week in the presence of three previously-reported MMC agents (carrageenan, Ficoll 70/400, and hyaluronic acid). We measured the resulting deposition of collagens and sulfated glycosaminoglycans using quantitative assays, as well as other matrix components by immunostaining. Both carrageenan and Ficoll promoted significantly more accumulation of total collagen content, sulfated glycosaminoglycan content, and fibronectin staining. Only Ficoll, however, also demonstrated a significant increase in collagen I staining. The results were more variable in 3D spheroid culture. We focused on Ficoll MMC matrices, which were isolated using the small molecule Raptinal to induce cancer cell apoptosis and matrix decellularization. The cancer cell-derived matrix promoted significantly faster migration of human astrocytes in a scratch wound assay, which may be explained by focal adhesion morphology and an increase in cellular metabolic activity. Ultimately, these data show MMC culture is a useful technique to generate cancer cell-derived matrices and study the effects on stromal cell migration related to wound repair.

Characterization of Proliferation Medium and Its Effect on Differentiation of Muscle Satellite Cells from Larimichthys crocea in Cultured Fish Meat Production

“…Among them, matrix is widely used to mimic the environment of the extracellular matrix (ECM), which assists in maintaining the optimal balance for a series of cell adjustment behaviors, including cell proliferation, migration and differentiation [31]. The ECM plays a major role in cell-cell and cell-matrix signaling during normal physiology and disease [32]. The results showed that there was no significant difference in the proliferation of PSCs between coated and uncoated vessels, and the effect of matrix was the best among the coating materials.…”

Decellularized tissues as platforms for digestive system cancer models

“…The addition of inert macromolecules to the cell culture medium helps mimic the crowded environment of living organisms by creating steric hindrance and electrostatic repulsion. This technique restricts the diffusion of cell-derived molecules such as procollagen and proteinases, which accelerates the deposition of ECM [ 84 ].…”