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S. Singhal, B. Bhatia, P. T. Khaw, G. A. Limb; Potential of MIO-M1 Cells to Differentiate Into Retinal Ganglion Cells (RGC) in vitro. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4065.
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© ARVO (1962-2015); The Authors (2016-present)
MIO-M1 (Moorfields Institute of Ophthalmology-Müller 1) cells, which exhibit Müller glial cell characteristics as judged by their gene expression and electrophysiological and phenotypic characteristics, also express markers of neural stem cells in vitro. This work investigated the potential of this cell line to differentiate along a retinal ganglion cell (RGC) fate.
We investigated the expression of transcription factors implicated in the developmental formation of RGC, including ATOH7, Brn-3b and differentiated RGC marker HuD (ELAVL4) in the MIO-M1 cells. We also examined the expression of cell signalling molecule Notch-1, a negative regulator of RGC differentiation, as well as the functional role of Notch-1 signalling in the differentiation of these cells in vitro.
MIO-M1 cells under normal culture conditions expressed low levels of ATOH7, Brn-3b and HuD. Expression of Brn-3b (a marker of committed RGC precursors) was significantly upregulated when the cells were cultured in the presence of specific growth factors and extracellular matrices. Endogenous Notch-1 signalling was detected in MIO-M1 cells and was down regulated when the cells were induced to differentiate. Inhibition of Notch-1 signalling using the gamma secretase inhibitor DAPT did not significantly improve expression of ATOH7 or Brn-3b in vitro.
MIO-M1 cells exhibit competence (ATOH7 expression) as well as commitment (Brn-3b expression) to differentiate into RGC. It is possible to induce these progenitors to differentiate into cells expressing RGC phenotype in vitro by altering the extracellular environment. When cells were induced to differentiate, endogenous Notch-1 signalling was downregulated. These results show that MIO-M1 cells exhibit some of the intrinsic programming elements of retinal progenitors during the developmental process of RGC formation, and suggest that MIO-M1 cells constitute a useful source of progenitors to form RGC in vitro.
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