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Xiaorong Li, Wenbo Li, Zhenyu Lu, Yunshan Zhang, Lijie Dong, Fei E. Wang, Rong Dong; Development Of Retinal Pigment Epithelium From Human Parthenogenetic Embryonic Stem Cells And Microrna Signature. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3949.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the potential of human parthenogenetic embryonic stem cells (hPESCs) to differentiate into retinal pigment epithelium (RPE) cells, and identify development-regulating microRNAs (miRNAs).
RPE cells were drived from hPESCs. The expression of markers, miRNA expression profiles and positional information of RPE ‘Signature’ miRNAs during differentiation were studied by using real-time RT-PCR, western blot and miRNA expression array at three time points. Human fetal RPE (hfRPE) cells were also analyzed. Then target genes of candidate miRNAs were validated.
RPE cells can be derived from hPESCs with an efficiency similar to human embryonic stem cells (hESCs). hPESCs-derived RPE cells exhibited similar morphology and pigmentation to hfRPE cells. However, the expression of markers (Oct4, Pax6, ZO-1, RPE65, MERTK) during retinal differentiation indicated that hPESCs-derived RPE cells were in an immature state. Most specific miRNAs played a role at some point in differentiation and maturation of RPE from hPESCs, with the exception of only two miRNAs (miR-204 and the miR-302s family). miR-204 showed an up-regulation, and miR-302 showed a down-regulation throughout the process. CTNNBIP1 and TGFBR2 were confirmed to be the target genes of miR-204 and miR-302 respectively, involved in mediating the activation of the Meis2/Pax6, Wnt/beta-catenin and TGFβ/Nodal pathways.
hPESCs can develop into RPE-like cells, which provides an additional promising source for RPE cells in cell therapy. miR-204, 302s and their targets are involved in regulating directed differentiation during the full course, which contribute to the search for a new method of improving the differentiation efficiency using miRNAs.
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