Abstract
Purpose: :
To investigate the regulatory effect of EGF on cell proliferation vs. differentiation in ARPE19 cells after different incubation periodes (24h vs. 48h) for obtaining ideal conditions for feasible rejuvenation and autologous transplantation of RPE cells.
Methods: :
To evaluate gene expression of RPE-specific differentiation and proliferation markers as well as transcriptional and translational activity of ß-Catenin signaling markers by FACS and RT-PCR after 24h and 48h EGF treatment.
Results: :
Our data showed a significant decrease of RPE 65, CRALBP and Cytokeratin 18 in ARPE-19 cells even after 24h EGF treatment. In addition an increase of Cyclin D1 expression and a significant decrease of GSK-3ß and ß-Catenin were equally observed after 24h and 48h EGF treatment. Cell cycle studies revealed an increase of ARPE cells in S-G2/M phase after 24h EGF treatment.
Conclusions: :
Our recently published data identified EGF as a potent initiator of RPE proliferation via activation of the ß-Catenin Signalling pathway after 48h treatment. Our new data demonstrate the induction of proliferation and upregulation of the ß-Catenin signaling pathway by EGF even after 24 hours incubation. As ideal cell culture conditions are essential to maintain RPE-specific phenotype, short incubation times enhance RPE cell quality for feasible rejuvenation and subsequent autologous transplantation of RPE cells.
Keywords: retinal pigment epithelium • age-related macular degeneration • aging