Construction of a multi-functional extracellular matrix protein that increases number of N1E-115 neuroblast cells
Makiko Nakamura1, Masayasu Mie, Hisakazu Mihara
1Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama 226-8501, Japan.
Abstract:
An artificially designed fusion protein, which was designed to have strong cell adhesive activity and an active functional unit that enhances neuronal differentiation of mouse N1E-115 neuroblast cells, was developed. In this study, a laminin-1-derived IKVAV sequence, which stimulates neurite outgrowth in conditions of serum deprivation, was engineered and incorporated into an elastin-derived structural unit. The designed fusion protein also had a cell-adhesive RGD sequence derived from fibronectin. The resultant fusion protein could adsorb efficiently onto hydrophobic culture surfaces and showed cell adhesion activity similar to laminin. N1E-115 cells grown on the fusion protein exhibited more cells with neurites than cells grown on laminin-1. These results indicated that the constructed protein could retain properties of incorporated functional peptides and could provide effective signal transport. The strategy of designing multi-functional fusion proteins has the possibility for supporting current tissue engineering techniques.
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