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A novel "sandwich" assay for quantifying chemo-regulated cell migration within 3-dimensional matrices: Wound healing cytokines exhibit distinct motogenic activities compared to the transmembrane assay

Research output: Contribution to journalArticlepeer-review

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Author(s)

S. L. Schor, I. R. Ellis, K. Harada, K. Motegi, Sandy Anderson, M. A. J. Chaplain, R. P. Keatch, A. M. Schor

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Abstract

The extracellular matrix profoundly affects cellular response to soluble motogens. In view of this critical aspect of matrix functionality, we have developed a novel assay to quantify chemo-regulated cell migration within biologically relevant 3-dimensional matrices. In this ?sandwich? assay, target cells are plated at the interface between an upper and lower matrix compartment, either in the presence of an isotropic (uniform) or anisotropic (gradient) spatial distribution of test motogen. Cell migration in response to the different conditions is ascertained by quantifying their subsequent disposition within the upper and lower matrix compartments. The objective of this study has been to compare the motogenic activities of platelet-derived growth factor (PDGF-AB) and transforming growth factor-beta isoforms (TGF-ß1, -ß2 and -ß3) in the sandwich assay and the commonly employed transmembrane assay. As previously reported, dermal fibroblasts exhibited a motogenic response to isotropic and anisotropic distributions of all tested cytokines in the transmembrane assay. In contrast, only PDGF-AB and TGF-ß3 were active in the sandwich assay, each eliciting directionally unbiased (symmetrical) migration into the upper and lower type I collagen matrices in response to an isotropic cytokine distribution and a directionally biased response to an anisotropic distribution. TGF-ß1 and -ß2 were completely devoid of motogenic activity. These results are consistent with the reported differential bioactivities of PDGF and TGF-ß3 compared to TGF-ß1 and -ß2 in animal models of wound healing and suggest that the sandwich assay provides a means of obtaining physiologically relevant data regarding chemo-regulated cell migration.
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Details

Original languageEnglish
Pages (from-to)287-300
Number of pages14
JournalCell Motility and the Cytoskeleton
Volume63
Issue number5
DOIs
Publication statusPublished - 2006

    Research areas

  • Cytokines, Wound healing, Chemotaxis, Collagen, TGF ?, PDGF

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