Novel extracellular matrix for cell sheet recovery using genetically engineered elastin‐like protein

Mie, Masayasu; Mizushima, Yasunori; Kobatake, Eiry

doi:10.1002/jbm.b.31019

Cited by 40 publications

(23 citation statements)

References 27 publications

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“…[9][10][11] In particular, elastin-like proteins (ELPs) modeled after human tropoelastin display soluble-insoluble phase transitions in response to changes in the environmental temperature, and these can be exploited for thermally controlled drug delivery or cellular engineering. 12,13 The ELP backbone is composed of the VPGXG pentapeptide, in which X is the guest position for any amino acid except proline. This pentapeptide is highly compatible with living cells.…”

Section: Introductionmentioning

confidence: 99%

Stimulation of fibroblasts and neuroblasts on a biomimetic extracellular matrix consisting of tandem repeats of the elastic VGVPG domain and RGD motif

Jeon

Park

Wei

et al. 2011

J Biomedical Materials Res

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Elastin-like proteins (ELPs) modeled after tropoelastin are favored in the development of biomimetic matrices due to their biocompatibility and the possibility to precisely control their environmental responsiveness, mechanical properties, and fate within the cells by recombinant DNA technology-mediated design at the gene level. However, a basic prerequisite in the use of ELPs as cell culture matrices is the presence of a biofunctionality that can induce adhesion-mediated signaling pathways. To activate fibronectin-integrin signaling events from a cell-matrix interface and direct cell survival and proliferation, we biosynthesized a modular ELP, represented as TGPG[VGRGD(VGVPG)₆]₂₀ WPC, consisting of alternating elastic (VGVPG)₆structural domains and cell-binding VGRGD motifs that are intended to emulate various aspects of extracellular matrix proteins. The inverse transition curves of [VGRGD(VGVPG)₆]₂₀ and (VGVPG)₁₄₀ overlapped with each other, indicating that one VGRGD sequence fused with six elastic pentapeptides did not disturb the thermal sensitivity of [VGRGD(VGVPG)₆]₂₀. The cell adhesion activity of [VGRGD(VGVPG)₆]₂₀ toward HEK293 fibroblasts and N2A neuroblasts was similar to that of native fibronectin. Upon contact with [VGRGD(VGVPG)₆]₂₀, the fibroblasts exhibited a flattened polygonal morphology, and the neuroblasts synthesized new DNA and proliferated. On the basis of these physiological changes, we concluded that RGD-functionalized ELP triggers the activation of signaling cascades within cells and can be used as an elastin-like matrix for mammalian cell culture.

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Section: Introductionmentioning

confidence: 99%

Stimulation of fibroblasts and neuroblasts on a biomimetic extracellular matrix consisting of tandem repeats of the elastic VGVPG domain and RGD motif

Jeon

Park

Wei

et al. 2011

J Biomedical Materials Res

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“…Furthermore, the ELPs potentially offer several advantages over PNIPAM such as biocompatibility and biodegradablility characters. Based on such interesting properties, ELPs have already been employed as useful molecular tools for biological applications such as protein purification, drug delivery and artificial ECM [17][18][19][20][21][22][23][24][25][26]. In most of these studies, genetically engineered ELPs with high molecular weights were used on the subjects.…”

Section: Introductionmentioning

confidence: 99%

Fabrication of Thermo-Responsive Molecular Layers from Self-Assembling Elastin-Like Oligopeptides Containing Cell-Binding Domain for Tissue Engineering

et al. 2015

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Novel thermo-responsive elastin-like oligopeptides containing cell-binding epitope (Arg-Gly-Asp-Ser sequence); arginine-glycine-aspartic acid-serine (RGDS)-elastin-like peptides (ELP) and RGDS-deg-ELP; were newly prepared as building blocks of self-assembled molecular layer for artificial extra cellular matrix. A detailed analysis of the conformation of the oligo(ELP)s in water and their self-assembling behavior onto hydrophobic surfaces were performed by using circular dichroism, Fourier transform infrared spectroscopy (FTIR), atomic force microscopy and water contact angle measurements. The experimental results revealed that both oligo(ELP)s self-assembled onto hydrophobic surfaces and formed molecular layers based on their thermo-responsive conformational change from hydrous random coil to dehydrated β-turn structure. Effective cell adhesion and spreading behaviors were observed on these self-assembled oligo(ELP) layers. In addition, attached cells were found to be recovered successfully as a cell-sheet by temperature-induced disassembly of oligo(ELP) layer. This achievement provides an important insight to construct novel oligopeptide-based nano-surfaces for the design of smart artificial extra-cellular matrix. OPEN ACCESSPolymers 2015, 7 135

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“…67 The cell sheets were obtained at 20°C after A549 cells were cultured on a culture dish coated with ELP containing GVGVP repeating units, His tag and RGD sequence. 46 Injectable biomaterials are of interest for the rapid and in vivo formation of load bearing scaffolds for tissue engineering. 35 This system is very attractive for tissue engineering because cells and growth factors can be homogeneously mixed with a liquid and injected into a defect site, followed by in situ formation of a hydrogel.…”

Section: Elastin-like Polymersmentioning

confidence: 99%

Application of Recombinant Fusion Proteins for Tissue Engineering

et al. 2010

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Extracellular matrix (ECM) plays important roles in tissue engineering because cellular growth and differentiation, in the two-dimensional cell culture as well as in the three-dimensional space of the developing organism, require ECM with which the cells can interact. Also, the development of new synthetic ECMs is very important because ECMs facilitate the localization and delivery of cells to the specific sites in the body. Therefore, the development of synthetic ECMs to replace the natural ECMs is increasingly essential and promising in tissue engineering. Recombinant genetic engineering method has enabled the synthesis of protein-based polymers with precisely controlled functionalities for the development of new synthetic ECMs. In this review, the design and construction of structure-based recombinant fusion proteins such as elastin-like polymers (ELPs) and silk-like polymers (SLPs), cell-bound growth factor-based recombinant fusion proteins such as basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF), hybrid system composed of recombinant protein and synthetic polymer, and E-cadherin-based fusion protein by recombinant genetic engineering were explained for application of the synthetic ECMs. Modulation of mechanical properties, stimuli-sensitivity, biodegradation and cell recognition can be achieved through precise control of sequence, length, hydrophobicity and cell binding domain by recombinant genetic engineering.

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Novel extracellular matrix for cell sheet recovery using genetically engineered elastin‐like protein

Cited by 40 publications

References 27 publications

Stimulation of fibroblasts and neuroblasts on a biomimetic extracellular matrix consisting of tandem repeats of the elastic VGVPG domain and RGD motif

Stimulation of fibroblasts and neuroblasts on a biomimetic extracellular matrix consisting of tandem repeats of the elastic VGVPG domain and RGD motif

Fabrication of Thermo-Responsive Molecular Layers from Self-Assembling Elastin-Like Oligopeptides Containing Cell-Binding Domain for Tissue Engineering

Application of Recombinant Fusion Proteins for Tissue Engineering

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