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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۷۱-۷۲
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عنوان انگلیسی Ps-119: Viability, Growth and Morphology of A Spontaneously Arising Human Retinal Pigment Epithelial Cell Line Those Express Stem/progenitor Cell Markers on Thin Layer of Alginate
چکیده انگلیسی مقاله Objective: Retinal pigment epithelium (RPE) is a highly polarized and specialized monolayer epithelium that placed at the interface between the photoreceptors of the neural retina and the vascular layer of the choroid. Almost all cells in the human body grow within organized 3D matrices. Alginate is naturally derived polysaccharide that, is an ideal bioMaterials for tissue engineering. culturing cells on alginate as a substrate provides a more in vivo-like environment. A spontaneously derived line of human RPE (hRPE) was found in our routine hRPE cultures. It revealed reasonable expression of stem/progenitor cell markers. We cultured the aforesaid hRPE line on alginate films and its apparent characteristics were investigated. Materials and Methods: 940 μl of 1% and 2% (v/w) alginate in deionized water were added in every well of 6 well micro plates for preparing a 1mm alginate film. Alginate substrate was gelatinized using 1X and 10X neurobasal cell culture medium (NCCM). hRPE cell line was isolated from neonatal human globes and cultured in DMEM/F12 supplement with 10% FBS. We characterized these cultures trough immunocytochemistry analysis for Oct4, Chx10, Pax6 and Ki67 markers. Cell were cultivated at a density of 200000 on each well of alginate coated 6 well micro plates. After 5 days RPE colonies were harvested and re-cultured on polystyrene substrates. Results: HRPE cell line survived for more than ten days on alginate film. Viability of the cultures was 99%. They grew very well on alginate films; total number of cells increased eleven times respect to controls. The cells formed big sticky colonies on alginate and exhibited bigger colonies on 2 %( v/w) alginate gelatinized with 10X (NCCM). When we harvested hRPE colonies and re-cultured them on polystyrene, cells did not switch their morphologic features and maintained their spheroidal structure. Conclusion: We found that alginate can support survival and growth of hRPE cell line and force them to organize in giant colony structures like spheroids. We are going to characterize hRPE cultures more precisely and investigate their differentiating potential into other retinal cells.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/923
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