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Cell transplantation to treat retinal degenerative diseases represents an option for the replacement of lost photoreceptor cells.  In vitro  expandable cells isolated from the developing mammalian retina have been suggested as a potential source for the generation of high numbers of donor photoreceptors. In this study we used standardized culture conditions based on the presence of the mitogens FGF-2 and EGF to generate high numbers of cells  in vitro  from the developing mouse retina. These presumptive ‘retinal stem cells’ (‘RSCs’) can be propagated as monolayer cultures over multiple passages, express markers of undifferentiated neural cells, and generate neuronal and glial cell types upon withdrawal of mitogens  in vitro  or following transplantation into the adult mouse retina. The proportion of neuronal differentiation can be significantly increased by stepwise removal of mitogens and inhibition of the notch signaling pathway. However, ‘RSCs’, by contrast to their primary counterparts  in vivo , i.e. retinal progenitor cells, loose the expression of retina-specific progenitor markers like  Rax  and  Chx10  after passaging and fail to differentiate into photoreceptors both  in vitro  or after intraretinal transplantation. Notably, ‘RSCs’ can be induced to differentiate into myelinating oligodendrocytes, a cell type not generated by primary retinal progenitor cells. Based on these findings we conclude that ‘RSCs’ expanded in high concentrations of FGF-2 and EGF loose their retinal identity and acquire features of  in vitro  expandable neural stem-like cells making them an inappropriate cell source for strategies aimed at replacing photoreceptor cells in the degenerated retina.

In Vitro Expanded Stem Cells from the Developing Retina Fail to Generate Photoreceptors but Differentiate into Myelinating Oligodendrocytes