The Use of 3-D Culture in Peptide Hydrogel for Analysis of Discoidin Domain Receptor 1-Collagen Interaction

• Published in: Cell adhesion & migration Vol. 1; no. 2; pp. 92 - 98
• Main Authors: Yoshida, Daizo, Teramoto, Akira
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 01.04.2007; Landes Bioscience
• Subjects: Binding; Biology; Bioscience; Calcium; Cancer; Cell; Cell Culture Techniques - methods; Cell Line, Tumor; Collagen - metabolism; Cycle; Discoidin Domain Receptors; Humans; Hydrogel, Polyethylene Glycol Dimethacrylate; Landes; Matrix Metalloproteinase 2 - metabolism; Matrix Metalloproteinase 9 - metabolism; Microscopy, Electron, Scanning; Neoplasm Invasiveness - pathology; Organogenesis; Peptides; Protein Binding; Proteins; Receptor Protein-Tyrosine Kinases - metabolism; Receptors, Mitogen - metabolism; Research Paper
• ISSN: 1933-6918; 1933-6926
• DOI: 10.4161/cam.1.2.4618

The aim of this study is to examine a novel drop culture model using a biologically inspired self-assembling peptide: hydrogel (RAD16-I, also called PuraMatrix), which produces a nanoscale environment similar to native extracellular matrix (ECM) for a cell line weakly adherent to a plastic surface during cell culture. Our work investigates quantitatively analyzing discoidin domain receptor (DDR) 1-mediated protein interactions between collagen type I and matrix metalloproteinase (MMP)-2 or -9, as well as cell invasion, using, as a scaffold, PuraMatrix, a novel peptide hydrogel. Results demonstrate that the dynamic cell culture technique produced a highly stable reharvesting of cells throughout the constructs with HP-75, human pituitary adenoma cell line when compared to the traditional seeding methods. Secretion of MMP via collagen type I was observed quantitatively in the supernatant (EC50; MMP-2, 50.4 ng/ml: MMP-9, 57.6 ng/ml). In PuraMatrix gel impregnated with 50 ng/ml of collagen type I, transfection of the vector encoding full-length DDR1 or siRNA targeting DDR1 up- or down-regulated respectively secretion of MMP-2 and -9, and cell invasion. Our results show that incorporation of this peptide with each ECM component provides a more permissive environment to elucidate ECM to cell signal interaction.