Protein-free cell culture on an artificial substrate with covalently immobilized insulin.

Ito, Yoshihiro; Zheng, Junchao; Imanishi, Yukio; Yonezawa, Kazuyoshi; Kasuga, Masato

doi:10.1073/pnas.93.8.3598

Cited by 128 publications

(83 citation statements)

References 27 publications

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“…These findings indicated that the immobilized insulin did not enhance cell adhesion but transduced a signal to the cells through phos- phorylation of tyrosine residues of signal proteins. Previously, we reported that immobilized insulin activated insulin receptors, insulin receptor substrate-1 and phosphatidylinositol-3-kinase for a longer period than free insulin [12]. Therefore, even after 8 h of culture, activated cells were observed on the insulin immobilized region.…”

Section: Resultsmentioning

confidence: 89%

“…The immobilized insulin activated the insulin receptor and downstream signaling proteins, and this activation persisted for longer periods than that with soluble insulin, probably explaining the greater mitogenic effect of the immobilized insulin [12]. In this study, signal transduction from the immobilized insulin was clearly visualized and the possibility of regulating cell functions through 'artificial juxtacrine stimulation' was examined using a micro-pattern-immobilized insulin.…”

Section: Introductionmentioning

confidence: 86%

“…Chinese hamster ovary cells overexpressing insulin receptors (CHO-T, 10 6 receptors per cell) [12] were subcultured in Ham's F-12 medium containing 10% (v/v) fetal bovine serum under 5% C0 2 in air at 37°C. After culture in the absence of serum for 2 days, cells were harvested by incubation at 37°C for 10 min with PBS containing 0.02% (w/v) EDTA, washed with Ham's F-12 medium twice, and suspended in Ham's F-12 medium (1 X10 5 cells/ml).…”

Section: Cell Culturementioning

confidence: 99%

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Patterned artificial juxtacrine stimulation of cells by covalently immobilized insulin

Ito¹,

Kondo²,

Chen³

et al. 1997

FEBS Letters

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Insulin was immobilized on prescribed areas of poly(ethylene terephthalate) film by photolithography. Chinese hamster ovary cells overexpressing human insulin receptors were cultured on the pattern-immobilized film. Pattern-immobilized insulin did not enhance cell adhesion but transduced a signal to the cells through phosphorylation of tyrosine residues of signal proteins. In addition, only cells on immobilized insulin grew in medium containing no serum. The enhancement of cell growth was considered to be a consequence of signal transduction. This microprocessing technique is useful for investigating the effect of immobilized biosignal molecules and for constructing new tissue engineering materials.

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Section: Resultsmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 86%

Section: Cell Culturementioning

confidence: 99%

See 1 more Smart Citation

Patterned artificial juxtacrine stimulation of cells by covalently immobilized insulin

Ito¹,

Kondo²,

Chen³

et al. 1997

FEBS Letters

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show abstract

“…[17] It was more than 20 years later when it was shown that low doses of insulin immobilized on polymers had higher mitogenic effect than free insulin. [18] Simultaneously, it was shown that epidermal GF (EGF) could be tethered to surfaces retaining its biological activity and with control of temporal and spatial availability. [19] However, besides these early successes it took another decade to investigate systematically the effect of GFs bound to surfaces.…”

Section: Introductionmentioning

confidence: 99%

Mechanotransduction and Growth Factor Signalling to Engineer Cellular Microenvironments

Cipitria

Salmerón-Sánchez

2017

Adv Healthcare Materials

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Engineering cellular microenvironments involves biochemical factors, the extracellular matrix (ECM) and the interaction with neighbouring cells. This progress report provides a critical overview of key studies that incorporate growth factor (GF) signalling and mechanotransduction into the design of advanced microenvironments. Materials systems have been developed for surface‐bound presentation of GFs, either covalently tethered or sequestered through physico‐chemical affinity to the matrix, as an alternative to soluble GFs. Furthermore, some materials contain both GF and integrin binding regions and thereby enable synergistic signalling between the two. Mechanotransduction refers to the ability of the cells to sense physical properties of the ECM and to transduce them into biochemical signals. Various aspects of the physics of the ECM, i.e. stiffness, geometry and ligand spacing, as well as time‐dependent properties, such as matrix stiffening, degradability, viscoelasticity, surface mobility as well as spatial patterns and gradients of physical cues are discussed. To conclude, various examples illustrate the potential for cooperative signalling of growth factors and the physical properties of the microenvironment for potential applications in regenerative medicine, cancer research and drug testing.

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“…The immobilized cytokines on the surface of a high molecular weight substrate continued to function without exhaustion of the cytokines. [14][15][16] Epidermal growth factor (EGF) among the various cytokines showed a ''juxtacrine stimulation'' effect to the target cells. 17,18 The present in vitro study was designed to examine the potential for early treatment of ACL injuries using an artificial juxtacrine cell culture model by a photo-immobilized EGF.…”

Section: Introductionmentioning

confidence: 99%

Proliferation of anterior cruciate ligament cells in vitro by photo‐immobilized epidermal growth factor

Woo

Kwon

Lee

et al. 2006

Journal Orthopaedic Research

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The purpose of this study was to explore the early treatment potential of anterior cruciate ligament (ACL) injuries using artificial juxtacrine stimulation by photo-immobilization of a growth factor. A photo-reactive epidermal growth factor complex (EGF-Az) was synthesized by conjugating EGF with N-(4-azidobenzoyloxy) succinimide followed by immobilization onto polystyrene culture plates using UV irradiation. ACL cells from human tissues (1 Â 10 5 cells, 100 ml/ well) were cultured as follows: control, no EGF; 50 ml native EGF; 50 ml EGF-Az immobilized; and 100 ml EGF-Az immobilized. The ACL cells were cultured long-term and evaluated for possible differences in their responses to EGF. An in vitro wound closure assay was developed to enable examination of cellular proliferation and migration. ACL cell proliferation was most evident in the photo-immobilized EGF culture group and was seen to increase in proportion to the amount of added EGF. In the in vitro wound closure assay, the lesioned area at 72 h after culture initiation was indistinguishable in the photo-immobilized cultures, but remained clearly visible in the controls. We conclude that photo-immobilized EGF induced rapid proliferation of ACL fibroblast cells by artificial juxtacrine stimulation and speculate that similar EGF immobilization onto bioabsorbable material (e.g., polyglycolic acid or polylactic acid) might contribute to a new therapy for the treatment of ACL injuries. ß

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Protein-free cell culture on an artificial substrate with covalently immobilized insulin.

Abstract: Insulin

Cited by 128 publications

References 27 publications

Patterned artificial juxtacrine stimulation of cells by covalently immobilized insulin

Patterned artificial juxtacrine stimulation of cells by covalently immobilized insulin

Mechanotransduction and Growth Factor Signalling to Engineer Cellular Microenvironments

Proliferation of anterior cruciate ligament cells in vitro by photo‐immobilized epidermal growth factor

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