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M. Yamaguchi, N. Ebihara, N. Shima, M. Kimoto, T. Funaki, S. Yokoo, A. Murakami, S. Yamagami; Laminin-5 Enhances Adhesion, Migration, and Proliferation Capacity of Cultured Human Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6423.
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To investigate the expression of laminin-5 (LM5) and its receptors on human corneal endothelial cells (HCEC) and whether exogenous recombinant human LM5 affects cultured HCEC adhesion, proliferation, and migration capacity.
Expression of LM5 and its receptors was examined in human donor corneas by immunohistochemistry and flow cytometry. Cultured HCECs were used for analysis of biological effects of LM5 on HCEC in serum-free condition. Changes of HCEC adhesion and proliferation by LM5 were evaluated by cell number counted with a Coulter counter. HCEC migration was assessed by the percentage wound closure quantified with an image processing and analysis software program in wound-healing assay.
Adult HCEC expresses LM5 receptor alpha3beta1 integrin, but not LM5. Significant higher number of cells adhered in an hour on recombinant LM5 (1.0 µg/ml) pre-coated dishes than those on non-coating dishes in cell adhesion assay. Cultured HCEC proliferation capacity was moderately promoted by coated LM5 (1.0 µg/ml) and soluble LM5 (20 ng/ml and 50 ng/ml) in proliferation assay. The percentage wound closure in the medium containing soluble LM5 after wounding was significantly higher than that in the control medium in wound-healing assay.
HCEC expresses LM5 receptor alpha3beta1 integrin. Exogenous LM5 has a capacity to promote adhesion, migration and moderate proliferation of cultured HCEC. LM5 can be a critical factor to facilitate HCEC culture and may contribute to practical use of tissue-engineered HCEC.
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