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Y. Arsenijevic, B. Angenieux, C. Kostic, D. Hornfeld, M. Tekaya, D.F. Schorderet, F.L. Munier; High Yield Of Photoreceptor–like Cells From Expanded Mouse Retinal Progenitor Cells. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):5410.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:In vitro generation of photoreceptors (PR) should be a promising tool to screen PR protective drugs and to provide an unlimited cell source for transplantation studies in animal models of retinal degeneration. We investigated whether expanded mouse retinal progenitor cells (RPCs) can produce PR in a defined medium. Methods:RPCs were isolated from postnatal DBA mice. The cells were grown in EGF and FGF–2 and were passed 3 to 17 times before inducing differentiation. For the induction of differentiation, 40'000 single cells were transferred onto an adhesive substrate. After different incubation times, the cells were either fixed or the RNA was extracted Results:Prior to differentiation, all RPCs express nestin filament and Pax6, two markers of precursor cells. They also show a high proliferative capacity, since RPCs from one retina can generate more than 7*1028 cells. Cells were induced to differentiate at different passages on an adhesive substrate. In a serum–free media without factors, no cells survived. In the presence of FGF–2, cells proliferated, forming flat cells and neuron–like cells. Immunocytochemistry analysis revealed that around 40% of the cells expressed b–tubulinIII, a neuronal marker, and presented an immature neuronal morphology (n=5). In view of these results, we first stimulated cells with FGF–2 and then changed the medium after five days to induce full differentiation and maturation. In this case, 40% of the cells expressed the photoreceptor marker recoverin (n=4). Such results were also obtained with cells from passage 18. In a preliminary experiment, we observed by RT–PCR that differentiated cells express the rod specific transcription factor Nrl, indicating that a population of cells was committed to the PR lineage. Further characterization of these PR–like cells is ongoing. Conclusions:Because RPCs of one retina can produce a generous amount of cells, equal to the number of blood cells of humankind and because they maintain the capacity to generate retinal cells even after extensive proliferation, we believe that RPCs are of great interest to study retinal neurogenesis and to provide cells for transplantation studies.
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