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Miguel Flores-Bellver, Sr., Violeta Sánchez-Vallejo, Raquel Álvarez-Nölting, Manuel Monleón Pradas, Jose Miguel Soria, María Miranda, Francisco Javier Romero; Characterization Of Isolated Retinal Progenitor Cells In Polymeric Scaffolds. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3938.
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Regenerative or cellular therapy has emerged in recent years as one of the most promising alternatives in the treatment of many diseases, including retinal degenerations such as retinitis pigmentosa or age related macular degeneration. A strategy to restore visual function in these patients is to replace the cells affected by retinal progenitor cells (RPC) that are predisposed to differentiate into neurons in the retina. On the other hand, obtaining and characterizing neurospheres from progenitor cells of different animal models of retinal disease can also be a way to study the mechanisms involved in these diseases. With this background, the objective of this work was to characterize and differentiate neurospheres derived from retina of mice into scaffolds or biomaterial thereby studying their integration and survival.
GFP+ Mice (C57BL/6J genetic background) were used in this study (n=5). The animal care and protocols were approved by the Ethics Committee of the institution and adjusted to the Spanish law concerning animal experiments.Progenitor cells were isolated from retina at postnatal day 1. We proceeded to culture these cells until day 7 and then seeded them on a biomaterial or scaffold. Biomaterials based on the hydrophobic polymer ethylacrylate, EA, and hydroxyethyl acrylate, p (EA-co-HEA 90-10) were prepared as polymeric scaffolds.Cells grown in the scaffold were characterized with the following markers: nestin, Ki-67, GFAP, MAP2, recoverin, TUJ-1 and NF200. We also proceeded to the differentiation of neurospheres to day 14 culturing them in the absence of growth factors to further characterize them with the same markers.
Neurospheres from progenitor cells express markers of retinal neurofilament and proliferation, like GFAP, TUJ1, Ki-67 and nestin. Differentiated cells express neuronal markers like TUJ-1 and photoreceptors markers (i.e. recoverin, rhodopsin).
Cells obtained from retinas have progenitor properties and neurogenic potential, providing potential sources of cells for transplantation through scaffolds.
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