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A. Kubota, K. Nakashima, K. Nishida, Y. Tano; Mammalian Müller Glial Cells Can Be Converted to a Neuronal Lineage by Combinatorial Method of Aggregation Culture With Neuronal Differentiation–Inducing Factors . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1068.
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It has been reported that in vivo, mammalian Müller glial cells could be converted to neuronal cells when stimulated by intravitreous injection of N–methyl–d–aspartate. However, its neuronal differentiation potential in vitro has yet to be confirmed. Here we report methods to induce neuronal differentiation of mammalian Müller glial cells in vitro.
Rat Müller glial cells were obtained from 2–week–old Long–Evans rats and cultured in the presence of serum. They were then passed onto non–adhesive coating culture dishes in the serum–free defined medium. After 5 days–culture, they were transferred to ornathine–laminin pre–coated chamberslides and further cultured for 7 days. Platelet–derived growth factor (PDGF) and varproic acid (VPA) were added to the culture either alone or simultaneously to stimulate neuronal differentiation.
Müller glial cells formed neurosphere–like aggregation on non–adhesive culture dishes. They became to express an immature neuronal marker ß–III tubulin when stimulated with PDGF and/or VPA.
Mammalian Müller glial cells could be converted to a neuronal lineage in vitro by a combinatorial method of aggregation culture with neuronal differentiation –inducing factors. These results imply that Müller glial cells could be a new source of neuronal cells for cell replacement therapy of such as retinitis pigmentosa.
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