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J. Ge, X. Sun, R. Jiang, Y. Zhang, Q. Gao, J. Zhuang, B. Huang, Y. Duan; E13.5 Retinal Precursor Cells Induce Bone Marrow Mesenchymal Stem Cells Towards Retinal Ganglion-Like Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5474.
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© ARVO (1962-2015); The Authors (2016-present)
To explore the possibility of targeting inducing of bone marrow mesenchymal stem cells (BMSCs) to differentiate towards retinal ganglion cells using neural retinal precursor cells (RPCs).
Mice RPCs from various developmental stages were harvested by traditional methods and co-cultured with BMSCs in transwell chamber. At different time points, RT-PCR, realtime qRT-PCR and immunofluorescence were performed in BMSCs to detect the expression of genes and proteins involved in retinal development.
Long-term culture of RPCs could be achieved with serum-free neuronal medium. BMSC displayed spindle shape before coculture and no expression of Pax6 and Brn3b could be detected by RT-PCR and immunofluorescence. After co-culturing with E13.5 RPCs for 3d, BMSCs displayed morphology changes from spindle shape to neurite-like cells. RT-PCR showed Rx, Pax6, Math5 and Brn3b, which are critical in retinal ganglion cells development, were expressed in BMSCs. qRT-PCR confirmed that expression of Pax6 and Brn3b increased by 439- and 70-fold, respectively, compared to BMSCs controls. Expression of Pax6 and Brn3b in BMSCs was further confirmed by immunofluorescence. Then the expression of Rx, Pax6, Math5 and Brn3b in BMSCs declined rapidly towards that of BMSCs control after continuing coculture with E13.5 RPCs for 7d, 14d and 21d. Furthermore, BMSCs displayed spindle shape again at 21d. When co-cultured with E17.5 RPCs, BMSCs displayed neurite-like morphology too. However, Rx, Pax6 and Brn3b could not be detected at any time points.
BMSCs can be specially induced to retinal ganglion-like cells by expressing of Math5 and Brn3b when co-culturing with E13.5 RPC for 3d. It offers a new insight into developmental genes modulating of special differentiation.
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