title: Characterization of a spontaneously generated murine retinal pigmented epithelium cell line; a model for in vitro experiments.
abstract: Retinal pigmented epithelium (RPE), the outermost layer of the retina, has a key role in maintaining retinal cells' functions. Severity of the culture of RPE cells has exerted many limitations to both in vitro and in vivo studies and its therapeutic applications. Therefore, establishment of RPE cell lines with high proliferative potential can considerably improve study of RPE cell biology. Here we report generation of a spontaneously immortalized murine RPE cell line in primary mouse RPE cell culture. Founded colonized cells were picked up and expression of RPE and retinal progenitor cells' (RPC) markers were studied using immunocytochemistry (ICC). Emerged cells cultured over 35 passages and population doubling times in different serum concentrations were calculated. We also investigated the ability of cells for becoming transfected by calcium-phosphate method and for becoming infected by adeno-associated virus serotype 2 (AAV2) using flow cytometry. Data showed that the cobblestone constituent cells expressed RPE65, cytokeratin and ZO1 and moreover several progenitor markers such as Pax6, Sox2, Nestin and Chx10. It revealed that, despite primary RPE cells, the newly emerged cells were easily transfectable and were highly infectable when compared with HEK293T cells. Our data indicated that the emerged mouse RPE cell line pretended RPC-like phenotype and also simultaneously expressed RPE markers. It would be a promising model for leading studies on RPE and RPC cells and substantially confirmed the great RPE plasticity and its invaluable potential in research studies.
authors.affiliation: Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Ehsan
authors.initials: E
authors.lastname: Ranaei Pirmardan
authors.affiliation: Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Zahra-Soheila
authors.initials: ZS
authors.lastname: Soheili
authors.affiliation: Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
authors.forename: Shahram
authors.initials: S
authors.lastname: Samiei
authors.affiliation: Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Hamid
authors.initials: H
authors.lastname: Ahmadieh
authors.affiliation: Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Seyed Javad
authors.initials: SJ
authors.lastname: Mowla
authors.affiliation: Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Razie
authors.initials: R
authors.lastname: Ezzati
authors.affiliation: Department of Molecular Medicine, Faculty of Advanced Technology, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Marzieh
authors.initials: M
authors.lastname: Naseri
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation; D048430:Cell Shape; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008954:Models, Biological; D055213:Retinal Pigment Epithelium / Q000166:cytology*; D013234:Stem Cells / Q000378:metabolism; D014162:Transfection
chemical_list: D015415:Biomarkers
keywords: Cell line; Infection; Retinal pigmented epithelium; Transfection