1. Characterization of Vitronectin Effect in 3D Ewing Sarcoma Models: A Digital Microscopic Analysis of Two Cell Lines.
- Author
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López-Carrasco, Amparo, Parra-Haro, Karina, Vieco-Martí, Isaac, Granados-Aparici, Sofía, Díaz-Martín, Juan, Salguero-Aranda, Carmen, Acevedo-León, Delia, de Álava, Enrique, Navarro, Samuel, and Noguera, Rosa
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VIRTUAL microscopy , *CANCER invasiveness , *CLUSTER analysis (Statistics) , *TUMORS in children , *RESEARCH funding , *ENZYME-linked immunosorbent assay , *GLYCOPROTEINS , *TUMOR markers , *CELL lines , *CELL culture , *SECRETION , *EWING'S sarcoma , *EXTRACELLULAR matrix , *THREE-dimensional printing , *COMPARATIVE studies , *METABOLOMICS , *PHENOTYPES ,BODY fluid examination - Abstract
Simple Summary: Three-dimensional (3D) models enable simplified and controlled in vitro replication of tumor microenvironment features, making them easier to study. In the current work, we developed 3D hydrogels based on gelatin plus silk fibroin with tunable mechanical properties, incorporating vitronectin (a glycoprotein related to tumor aggressiveness) into the scaffolds. Digital analysis was applied to monitor Ewing sarcoma cell line growth and morphology in response to the 3D model composition and different culture times, opening the possibility of its use to study other proteins of interest and tumor types. Our findings highlight the heterogeneity of cell line responses to their microenvironment, underscoring the enormous potential of these hydrogels as mechanodrug-testing platforms and for studying cell protein secretion as an initial step in liquid biopsy biomarker research. Ewing sarcoma (ES) is an aggressive bone and soft-tissue pediatric cancer. High vitronectin (VN) expression has been associated with poor prognosis in other cancers, and we aimed to determine the utility of this extracellular matrix glycoprotein as a biomarker of aggressiveness in ES. Silk fibroin plus gelatin–tyramine hydrogels (HGs) were fabricated with and without cross-linked VN and cultivated with A673 and PDX73 ES cell lines for two and three weeks. VN secretion to culture media was assessed using ELISA. Morphometric analysis was applied for phenotypic characterization. VN release to culture media was higher in 3D models than in monolayer cultures, and intracellular, intercellular, and pericluster presence was also observed. A673-HGs showed lower density of clusters but a proportion of larger clusters than PDX73-HGs, which presented low cluster circularity. The cluster density of A673-HGs without added VN was higher than with added VN and slightly lower in the case of PDX73-HGs. Furthermore, a culture time of three weeks provided no benefits in cluster growth compared to two weeks, especially in A673-HGs. These advances in 3D modeling and digital quantification pave the way for future studies in ES and other cancers to deepen understanding about intra- and intercellular heterogeneity and anti-adhesion VN therapies. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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