title: Isolation, Characterization, and Establishment of Spontaneously Immortalized Cell Line HRPE-2S With Stem Cell Properties.
abstract: The retinal pigment epithelium is a monolayer of highly specialized pigmented cells located between the neural retina and the Bruch's membrane of the choroid. RPE cells play a crucial role in the maintenance and function of the underlying photoreceptors. This study introduces a spontaneously arising human retinal pigment epithelial cell line, HRPE-2S, which was isolated from primary RPE cell culture of 2 days old male donor. We characterized morphology and functional properties of the new cell line. The immortalized cell line was maintained in culture for more than 70 passages and 240 divisions. The average doubling time of the cells was approximately 22 h and got freezed at 26th passage. The cell line expressed RPE-specific markers RPE65 and cell junction protein ZO1 as an epithelial cell marker. It also expressed CHX10, PAX6, Nestin, SOX2 as stem and retinal progenitor cell markers. Ki67 as a marker of cell proliferation was expressed in all HRPE-2S cells. It represented typical epithelial cobblestone morphology and did not phenotypically change through several passages. Stem cell-like aggregations (neurospheres) were observed in SEM microscopy. The cells represented high mitotic index. They could be viable under hypoxic conditions and serum deprivation. According to functional studies, the cell line exhibited stem cell-like behaviors with particular emphasis on its self-renewal capacity. LDH isoenzymes expression pattern confirmed the same cellular source for both of the HRPE-2S cells and primary RPE cells. Characteristics of HRPE-2S cells promise it as an in vitro model for RPE stem cell-based researches. J. Cell. Physiol. 232: 2626-2640, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Hoda
authors.initials: H
authors.lastname: Shams Najafabadi
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: Ocular Tissue Engineering 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: 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: Maryam
authors.initials: M
authors.lastname: Masoumi
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D015687:Cell Hypoxia; D002461:Cell Line, Transformed; D002465:Cell Movement; D049109:Cell Proliferation; D000066673:Cell Self Renewal; D002469:Cell Separation / Q000379:methods*; D048430:Cell Shape; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D008855:Microscopy, Electron, Scanning; D008940:Mitotic Index; D064231:Nestin / Q000378:metabolism; D000071841:PAX6 Transcription Factor / Q000378:metabolism; D010641:Phenotype; D055213:Retinal Pigment Epithelium / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D018874:Spheroids, Cellular; D013234:Stem Cells / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000378:metabolism; D019745:cis-trans-Isomerases / Q000378:metabolism; D000099178:Retinoid Isomerohydrolase
chemical_list: D015415:Biomarkers; D018398:Homeodomain Proteins; D019394:Ki-67 Antigen; D064231:Nestin; D000071841:PAX6 Transcription Factor; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; D062826:Zonula Occludens-1 Protein; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase; C577484:NES protein, human; C000605730:PAX6 protein, human; C527782:SOX2 protein, human; C569042:TJP1 protein, human; C490289:VSX2 protein, human