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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۷۱-۷۱
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عنوان انگلیسی Ps-118: Expression of Stem and Progenitor Cell bMarkers in A Spontaneously Arising Human Retinal Pigment Epithelial Cell Line
چکیده انگلیسی مقاله Objective: RPE is a monolayer of cells located between vessels of the choriocapillaris and light-sensitive outer segments of the photoreceptors. During the embryonic development of the eye, retinal stem cells in the optic vesicle give rise to multipotent progenitor cells that generate all the neurons and the Muller glia of the mature retina.This study describes morphology, viability, growth and characterization of a spontaneously arising human RPE (hRPE) cell line that have been found in our routine hRPE cell culture. Materials and Methods: RPE was isolated from neonatal human globes and cultured in DMEM/F12 supplement with 10% FBS. Cultures were continuously studied using phase contrast microscopy. After the 9th passage cultured cells were characterized trough immunocytochemistry analysis for Oct4, Chx10, and Pax6 and Ki67 markers. Results: The first few passages of the cultures were purely RPE cells, those revealed usual morphological features of the pigmented epithelium. Then suddenly alternative variant of cells appeared They made sizeable holes between RPE cells’ monolayer and push surround of this holes. They finally push all of the culture vessels and RPE cells’ features totally disappeared. The new founded cells were smaller than RPE cells and showed an appreciable tendency to form giant macroscopic colonies. After sequential passages they got smaller than before with a high nucleocytoplasmic ratio, representing thin layer of cytoplasm around the nucleus. The aforementioned cultures were passaged for 13 times until now. They grew faster than native RPE cells and rapidly covered culture surface .When we assessed for viability they showed more than 99% viable. Immunocytochemistry test revealed that 100% of the obtained cells expressed Chx10 whereas just 4% were pax6+ .60% of the studied nuclei were Ki67+ and 50% expressed Oct4. Conclusion: Our human RPE cultures gave rise to a cell line of hRPE that exhibited retinal cell stemness markers.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/922
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