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S. He, S. Kase, J. Xu, M. Kitamura, C. Kim, S. Ryan, D. Hinton; RPE Wound Healing Induces Upregulation of Laminin 5. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1840.
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© ARVO (1962-2015); The Authors (2016-present)
Extracellular matrix (ECM) production plays an important role in the wound healing response. The purpose of this study was to investigate the expression of Laminin 5 in a wound healing model using human retinal pigment epithelial cells (RPE) and the effects of laminin 5 on RPE cell function.
Confluent cultures of early passage human fetal RPE cells were scratched with a pipette tip and Laminin 5 expression in the wounded area was demonstrated by immunofluorescent staining and western blot at different time points after wounding. Laminin 5 mRNA expression in RPE cells was revealed by RT-PCR. Laminin 5 effects on RPE cell attachment and proliferation were evaluated using a modified MTT assay.
Cultured human RPE cells express Laminin 5 mRNA and protein. Wounding of the RPE monolayer resulted in an abundant increase of Laminin 5 at 16-24 hours after scratch as demonstrated by immunofluorescent staining and western blot. RPE cell attachment was reduced by 50% at 5 minutes and 30% at 10 minutes (P<0.01); RPE proliferation was inhibited by 30% at the concentration of 1 ug/ml Laminin 5 (p<0.05).
RPE cell attachment and proliferation were significantly inhibited by treatment with recombinant Laminin 5.Conclusions: Laminin 5 is upregulated in response to wound healing in RPE cells. Recombinant Laminin 5 inhibits RPE attachment and proliferation.
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