Construction of a multi‐functional extracellular matrix protein that increases number of N1E‐115 neuroblast cells having neurites

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.

Hydrogel scaffolds composed of genetically synthesized self-assembling peptides for three-dimensional cell culture

“…[2][3][4][5] Artificial ECM proteins have been designed and genetically synthesized in our laboratory. [6][7][8][9][10] Our strategy for the design of artificial ECM proteins relies on the combination of structural peptides and active functional peptides that control cellular functions. One of the ECM proteins previously constructed in our lab, named ERE, is composed of 12 repeats of the elastin-derived APGVGV peptide motif as the stable structural peptide, with the cell adhesive RGD peptide motif as the active functional peptide.…”

“…EREI, which consists of ERE fused with the IKVAV peptide, not only exhibits cell adhesive activity but also promotes angiogenesis and neural differentiation as a result of the combination of IKVAV and RGD. [8][9][10][11][12] In our genetic engineering-based designs, multi-functional ECMs composed not only of peptide motifs but also of growth factors were constructed. Epidermal growth factor fused to the ERE protein was found to exhibit cell growth activity via the epidermal growth factor moiety.…”

“…7 Our genetically designed ECM proteins based on ERE demonstrated applicability as biomaterials. [6][7][8][9][10] However, covalent cross-linking is required to form a hydrogel structure in order to apply our designed ECM proteins to three-dimensional (3-D) culture. 13 The use of cross-linking reagents can result in the loss of the functional peptide's activity.…”

Biomatrices and biomaterials for future developments of bioprinting and biofabrication

“…We consider that it is important to explore additional suitable hydrogel materials in future studies for advances in bioprinting and biofabrication technologies. Recent biochemical and nano-biotechnology techniques have introduced modified biomatrix materials, such as RGDmodified alginate hydrogel, and ECM proteins immobilized with growth factors and biofunctional peptide domains [36][37][38][39][40][41][42]. Indeed, exploratory research projects for more suitable and effective materials for use with bioprinting and biofabrication procedures are underway.…”

Construction of a tissue-specific transcription factor-tethered extracellular matrix protein via coiled-coil helix formation

“…6 A combination of functional units has showed synergistic effects. 7,8 To improve the functional capacities of these artificial ECMs, growth factors were attached to artificial ECMs, both covalently and non-covalently. [9][10][11][12] For the construction of biomaterials, combinations of ECMs and growth factors have been well studied owing to the fact that some growth factors are bound to ECMs under physiological conditions.…”