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A spontaneously arising RPE cell line from human RPE cell primary cultures

Session Details

Session Title: Quick Fire Free Paper Session 03

Session Date/Time: Sunday 29/09/2013 | 08:30-09:30

Paper Time: 09:40

Venue: Hall C (Level 1)

First Author: H.Ahmadieh IRAN

Co Author(s):    F. Javidi   Z. Soheili   S. Samiei        

Abstract Details

Purpose:

Retinal pigment epithelium (RPE) is composed of pigmented, single-layered, polarized cells located between the neural retina and the vascular choriocapillaris. While the neural retinal progenitor cells proliferate further to form the multi-layered retina, RPE becomes post mitotic and differentiates. Placing isolated RPE into a variety of defined culture conditions releases their normal quiescence. This study describes the development and characterization of a spontaneously arising human RPE cell line.

Setting:

National Institute of Genetic Engineering and Biotechnology

Methods:

RPE cells were isolated from neonatal human globes and cultured in Dulbecco's Modified Eagle's Medium (DMEM) /F12 supplement with 20% fetal bovine serum (FBS) and then consecutively sub-cultured in DMEM/F12 supplement with 10%FBS. Isolated RPE cells were characterized by RPE65 and cytokeratin 8/18.

Results:

Confluent cells were serially passaged 15 times. They displayed undifferentiated cells characteristics like high nucleoplasmic ratio and highly proliferative potential. Immunocytochemistry analysis revealed that all of cells express Ki67 marker and yet they share the RPE epithelial character and express epithelial cells cytokeratin. When we searched for Pax6, a reasonable number of cells showed positive signal for the marker.

Conclusions:

These findings reveal a previously unappreciated plasticity of RPE in humans. These observations indicate that RPE may have the capacity for self-repair under the correct circumstances, and other remarkable capacities that are relevant to human diseases in which RPE changes, proliferates, or degenerates, such as RP and AMD.

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