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Ana Fernandez-Gonzalez, Carlos Alonso-Ron, Manuel Chacon, Alvaro Meana, Jesus Merayo-Lloves, Carlos Belmonte; Influence Of Hyaluronan On Corneal Epithelium Cell Proliferation And Migration ‘In Vitro’. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3578.
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© ARVO (1962-2015); The Authors (2016-present)
To analyze the influence of sodium hyaluronan (HA) of different molecular weight, on the proliferation and migration rates of human epithelium cells in culture
Corneal epithelium cells obtained from limbal rings of human donors were expanded over irradiated mouse dermal fibroblasts (3T3) in the presence of serum, adenine, insulin, cholera toxin, triiodotironin and hydrocortisone. After 3 days, epidermal growth factor was added to the culture medium. 10-15 days later, cells were platted over the different substrates. In another set of experiments, SV40-transformed human corneal epithelial cells were cultured in the presence of serum. Limbal and transformed cells were serum-starved overnight before assay. Cell proliferation was measured using the DNA-stain based "CyQUANT™" (Invitrogen) kit. For cell migration determination, Boyden-Chamber based kits (Millipore) were used. Cell viability was assessed using a Wst-1 based kit (Roche). HA solutions of different molecular weights (50-5200 kDa) were added to the culture medium
HA had a variable effect on human corneal epithelium cells DNA synthesis depending on its MW. MW also determines the inhibition of migration of SV40-transformed human corneal epithelium cells by HA
HA promotes corneal epithelial wound healing by stimulating the migration and proliferation of the corneal epithelium. Our preliminary data suggest that the MW of HA plays a role in the corneal epithelium cell responses during wound healing
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