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Open Access Research article

Enhanced generation of retinal progenitor cells from human retinal pigment epithelial cells induced by amniotic fluid

Fatemeh Sanie-Jahromi1, Hamid Ahmadieh2*, Zahra-Soheila Soheili1, Maliheh Davari1, Shima Ghaderi1, Mozhgan Rezaei Kanavi2, Shahram Samiei3, Abdolkhalegh Deezagi1, Jalil Pakravesh4 and Abouzar Bagheri1

Author Affiliations

1 National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

2 Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

3 Iranian Blood Transfusion Organization Research Center, Tehran, Iran

4 Department of Obstetrics and Gynecology, Aban General Hospital, Tehran, Iran

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BMC Research Notes 2012, 5:182  doi:10.1186/1756-0500-5-182

Published: 10 April 2012

Abstract

Background

Retinal progenitor cells are a convenient source of cell replacement therapy in retinal degenerative disorders. The purpose of this study was to evaluate the expression patterns of the homeobox genes PAX6 and CHX10 (retinal progenitor markers) during treatment of human retinal pigment epithelium (RPE) cells with amniotic fluid (AF), RPE cells harvested from neonatal cadaver globes were cultured in a mixture of DMEM and Ham's F12 supplemented with 10% FBS. At different passages, cells were trypsinized and co-cultured with 30% AF obtained from normal fetuses of 1416 weeks gestational age.

Results

Compared to FBS-treated controls, AF-treated cultures exhibited special morphological changes in culture, including appearance of spheroid colonies, improved initial cell adhesion and ordered cell alignment. Cell proliferation assays indicated a remarkable increase in the proliferation rate of RPE cells cultivated in 30% AF-supplemented medium, compared with those grown in the absence of AF. Immunocytochemical analyses exhibited nuclear localization of retinal progenitor markers at a ratio of 33% and 27% for CHX10 and PAX6, respectively. This indicated a 3-fold increase in retinal progenitor markers in AF-treated cultures compared to FBS-treated controls. Real-time PCR data of retinal progenitor genes (PAX6, CHX10 and VSX-1) confirmed these results and demonstrated AF's capacity for promoting retinal progenitor cell generation.

Conclusion

Taken together, the results suggest that AF significantly promotes the rate of retinal progenitor cell generation, indicating that AF can be used as an enriched supplement for serum-free media used for the in vitro propagation of human progenitor cells.

Keywords:
Retinal progenitor cells; Amniotic fluid; Age related macular degeneration (AMD); Cellular therapy; Serum-free