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Ming-Feng Wu, Tanja Stachon, jiong wang, Xuefei Song, Sarach Colanesi, Achim Langenbucher, Berthold Seitz, Nora Szentmary; Effect of human autologous serum and fetal bovine serum on human corneal epithelial cell viability, migration and proliferation in vitro. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5620.
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© ARVO (1962-2015); The Authors (2016-present)
Autologous serum (AS) eye drops offer a potential treatment alternative for non-healing corneal epithelial defects in clinical practice. In corneal epithelial cell cultures, fetal bovine serum (FBS) is often used to support the growth of the cells. The dose-dependent effect of AS and FBS on viability, migration and proliferation of human corneal epithelial cells (HCECs) have not been specified yet. The purpose of this study was to analyse the concentration-dependent effects of AS and FBS on HCEC viability, migration and proliferation, in vitro.
First, AS was prepared from 13 patients according to the regulations of the LIONS Cornea Bank Saar-Lor-Lux, Trier/Westpfalz. HCECs were firstly cultured in DMEM/F12 with 5% FBS, 0.5% DMSO, 10 ng/mL human epidermal growth factor, 1% insulin-transferrin-selenium, then were incubated in serum media which was consisting of DMEM/F12 supplemented by 5%, 10%, 15% or 30% AS or FBS for 24 hours. Thereafter, HCEC viability was analysed using Cell Proliferation Kit XTT, HCEC migration using wound healing assay, HCEC proliferation by the cell proliferation ELISA BrdU (colorimetric) kit. Statistical analysis was performed using a linear mixed model in the framework of a Generalized Estimating Equations (GEE) approach to analyse the effect of AS and FBS using IBM-SPSS version 22.
Viability and migration of HCEC was significantly higher using AS compared to FBS (p=0.01 and p<0.01), but proliferation did not differ significantly between both groups (p=0.79). Viability of HCEC was dependent on the concentration of both FBS and AS (p=0.03), but migration and proliferatin was not (p=0.62 and p=0.28). Migration and proliferation of HCEC (p<0.01 for both) using AS or FBS were correlated, without correlation in HCEC viability (p=0.48).
Viability and migration of HCEC is better using AS compared to PBS, without difference in HCEC proliferation. Only viability of HCEC is dependent on concentrations of AS or FBS.
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