Development of Retinal Pigment Epithelium from Human Parthenogenetic Embryonic Stem Cells and MicroRNA Signature
Development of Retinal Pigment Epithelium from Human Parthenogenetic Embryonic Stem Cells and MicroRNA Signature
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人类孤雌胚胎干细胞视网膜色素上皮的发育和 MicroRNA 特征
DOI：
10.1167/iovs.12-8303
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发表时间：
2012-08-01
影响因子：
4.4
通讯作者：
Li, Xiao-Rong
中科院分区：
文献类型：
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作者：
Li, Wen-Bo;Zhang, Yun-Shan;Li, Xiao-Rong
目的.我们研究了人单性生殖胚胎干细胞（hPESCs）向RPE细胞分化的潜能，并鉴定了发育调控microRNAs（miRNAs）。RPE细胞来源于hPESC。采用免疫细胞化学、实时荧光定量RT-PCR和miRNA表达阵列技术，检测3个时间点细胞分化过程中标志物的表达和miRNA表达谱。还分析了人胎儿RPE（hfRPE）细胞。然后验证候选miRNAs的靶基因。hPESC衍生的RPE细胞表现出与hfRPE细胞相似的形态和色素沉着。分化过程中标记物的表达表明hPESC衍生的RPE细胞是未成熟的。除了两种miRNA（miR-204和miR-302家族）之外，大多数特异性miRNA在来自hPESC的RPE分化和成熟过程中的某个时间点起作用。在整个过程中，miR-204上调，miR-302下调。随后，与对照组相比，在miR-204过表达组和miR-302抑制组中色素簇和RPE标签基因表达显著增加。CTNNBIP 1和TGFBR 2分别是miR-204和miR-302的靶基因。hPESC可以发育成RPE样细胞，因此可以成为细胞治疗中RPE细胞的额外有前途的来源。miR-204、miR-302及其靶点参与调控整个过程中的定向分化，从而有助于寻找利用miRNA提高分化效率的新方法。（Invest Ophthalmol维斯科学。2012;53：5334-5343）DOI：10.1167/iovs.12-8303
PURPOSE. We investigated the potential of human parthenogenetic embryonic stem cells (hPESCs) to differentiate into RPE cells, and identified development-regulating microRNAs (miRNAs).METHODS. 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.RESULTS. 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.CONCLUSIONS. 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. (Invest Ophthalmol Vis Sci. 2012;53:5334-5343) DOI: 10.1167/iovs.12-8303