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Wen-Bo Li, Yun-Shan Zhang, Zhen-Yu Lu, Li-Jie Dong, Fei E. Wang, Rong Dong, Xiao-Rong Li; Development of Retinal Pigment Epithelium from Human Parthenogenetic Embryonic Stem Cells and MicroRNA Signature. Invest. Ophthalmol. Vis. Sci. 2012;53(9):5334-5343. doi: 10.1167/iovs.12-8303.
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© ARVO (1962-2015); The Authors (2016-present)
We investigated the potential of human parthenogenetic embryonic stem cells (hPESCs) to differentiate into RPE cells, and identified development-regulating microRNAs (miRNAs).
RPE cells were derived from hPESCs. The expression of markers and miRNA expression profiles during differentiation were studied by immunocytochemistry, real-time RT-PCR, and miRNA expression array at three time points. Human fetal RPE (hfRPE) cells also were analyzed. The target genes of candidate miRNAs then were validated.
hPESC-derived RPE cells exhibited similar morphology and pigmentation to hfRPE cells. The expression of markers during differentiation indicated that the hPESC-derived RPE cells were immature. Most specific miRNAs had a role at some time point during the differentiation and maturation of RPE from hPESCs, except for two miRNAs (miR-204 and the miR-302 family). The miR-204 was upregulated and miR-302 was down-regulated throughout the process. Subsequently, pigmented clusters and RPE signature gene expression increased significantly in the miR-204 overexpression group and miR-302 inhibition group compared to the control groups. CTNNBIP1 and TGFBR2 were confirmed to be the target genes of miR-204 and miR-302, respectively.
hPESCs can develop into RPE-like cells and, thus, can be additional promising sources of RPE cells in cell therapy. The miR-204, miR-302s, and their targets are involved in regulating directed differentiation during the full course, thereby contributing to the search for a new method of improving differentiation efficiency using miRNAs.
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