The traditional 2d culture has been proved inferior to reproduce the subtle interaction between cell-to-cell and cell-to-extracellular matrix (ECM) in tumor microenvironment (TME) and collagen in ECM contributes to various malignancies of tumors. Hence, the 3d model contained with collagen may overcome the shortcomings of 2d culture. In this study, the in vitro TME mimicking matrix was prepared by coupling porcine liver-derived collagen (COL) and the dialdehyde group of partially oxidized alginate (OA), namely OA-COL, and the 3d OA-COL droplets were polymerized by divalent calcium ions. In the 3d OA-COL droplets, cancer cells displayed vigorous proliferation, and the cells grew in clusters and formed a unique spindle like clone. Quantitative analysis proved that various gene transcription and protein expression were up-regulated for the cells in the 3d OA-COL droplets, including F-actin reassembling, focal adhesion, pseudopodia formation, and the proteins involved in epithelial-to-mesenchymal transition (EMT). The 3d OA-COL droplets induced the cells with strengthened polarity, invasiveness, higher IC50, and manifested stronger tumorigenicity in vivo. The fabricated 3d OA-COL droplets reproduced a variety of TME parameters, constructed an in vitro model similar to the TME in vivo, and it may facilitate many investigations in cell biology and tumor biology.
Keywords: Collagen; Mimicking; Oxidized alginate; Porcine liver; Three-dimensional (3d) culture; Tumor microenvironment (TME).
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