Brain Cancer Cell-Derived Matrices and Effects on Astrocyte Migration.
Brain Cancer Cell-Derived Matrices and Effects on Astrocyte Migration.
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脑癌细胞衍生基质及其对星形胶质细胞迁移的影响。
DOI：
10.1159/000522609
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Cornelison,RChase
中科院分区：
文献类型：
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作者：
Louisthelmy,Rebecca;Burke,BrycenM;Cornelison,RChase
细胞衍生基质是研究不同细胞类型的细胞外基质（ECM）和测试对细胞迁移或伤口修复的影响的有用工具。这些基质通常使用具有抗坏血酸的延长培养物产生以促进ECM产生。将这种技术应用于癌细胞培养可以推进癌症ECM及其对肿瘤微环境的招募和训练的影响的研究，但抗坏血酸对癌细胞具有潜在的细胞毒性。基于排除体积原理，还可以添加大分子拥挤（MMC）剂以增加基质沉积。我们报告使用MMC单独作为一种有效的策略，以产生脑癌细胞衍生的矩阵下游分析和细胞迁移的研究。我们将小鼠胶质母细胞瘤细胞系GL 261在三种先前报道的MMC剂（角叉菜胶、Ficoll 70/400和透明质酸）存在下培养1周。我们使用定量分析测量了胶原蛋白和硫酸化糖胺聚糖的沉积，以及通过免疫染色测量了其他基质成分。角叉菜胶和Ficoll均促进总胶原含量、硫酸化糖胺聚糖含量和纤维连接蛋白染色显著增加。然而，仅Ficoll也显示胶原蛋白I染色显著增加。结果在3D球状体培养中变化更大。我们专注于Ficoll MMC基质，其使用小分子Raptinal分离以诱导癌细胞凋亡和基质脱细胞化。在划痕试验中，癌细胞衍生的基质促进人星形胶质细胞显著更快的迁移，这可以通过粘着斑形态学和细胞代谢活性的增加来解释。最后，这些数据表明MMC培养是一种有用的技术，用于产生癌细胞衍生的基质，并研究对与伤口修复相关的基质细胞迁移的影响。
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 (MMC) agents can also be added to increase matrix deposition based on the excluded volume principle. We report the use of 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.