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Dynamic microRNA Profiles of Hepatic Differentiated Human Umbilical Cord Lining-Derived Mesenchymal Stem Cells

机译:肝分化的人类脐带衬砌间充质干细胞的动态微小RNa谱

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摘要

Despite the extensive hepatic differentiation potential of human umbilical cord lining-derived mesenchymal stem cells (hUC-MSC), little is known about the molecular mechanisms of hUC-MSC differentiation. At the post-transcriptional level, microRNAs are key players in the control of cell fate determination during differentiation. In this study, we aimed to identify microRNAs involved in the hepatic differentiation of hUC-MSCs. After successfully isolating hUC- MSCs, we induced hepatocyte formation in vitro with growth factors. After 26 days of induction, hUC-MSCs could express hepatocyte-specific genes, synthesize urea and glycogen and uptake low-density lipoprotein. Cellular total RNA from hUC-MSCs and hepatic differentiated hUC-MSCs was collected at 7 time points, including 2 days, 6 days, 10 days, 14 days, 22 days and 26 days, for microRNA microarray analysis. Dynamic microRNA profiles were identified that did not overlap or only partially overlapped with microRNAs reported to be involved in human liver development, hepatocyte regeneration or hepatic differentiation of liver-derived progenitor cells. A total of 61 microRNAs among 1205 human and 144 human viral microRNAs displayed consistent changes and were altered at least 2-fold between hUC-MSCs and hepatic differentiated hUC-MSCs. Among these microRNAs, 25 were over-expressed; this over-expression occurred either gradually or increased sharply and was maintained at a high level. A total of 36 microRNAs were under-expressed, with an expression pattern similar to that of the over-expressed microRNAs. The expression of the altered expressed microRNAs was also confirmed by quantitative reverse-transcription polymerase chain reaction. We also found that microRNAs involved in hepatic differentiation were not enriched in hepatocyte or hepatocellular carcinoma cells and can potentially target liver-enriched transcription factors and genes. The elucidation of the microRNA profile during the hepatic differentiation of hUC-MSCs provides the basis for clarifying the role of microRNAs in hUC-MSC hepatic differentiation and specific microRNA selection for the conversion of hUC-MSCs to hepatocytes.
机译:尽管人类脐带衬里的间充质干细胞(hUC-MSC)具有广泛的肝分化潜能,但对hUC-MSC分化的分子机制知之甚少。在转录后水平,microRNA是分化过程中控制细胞命运决定的关键因素。在这项研究中,我们旨在鉴定与hUC-MSC肝分化有关的microRNA。成功分离hUC-MSC后,我们在体外用生长因子诱导了肝细胞的形成。诱导26天后,hUC-MSCs可以表达肝细胞特异性基因,合成尿素和糖原并摄取低密度脂蛋白。在7个时间点(包括2天,6天,10天,14天,22天和26天)收集来自hUC-MSC和肝分化的hUC-MSC的细胞总RNA,用于微RNA微阵列分析。确定了动态microRNA谱,该谱与报道的与人类肝脏发育,肝细胞再生或肝祖细胞的肝分化有关的microRNA不重叠或仅部分重叠。在1205个人和144个人的病毒微RNA中,共有61个微RNA表现出一致的变化,并且在hUC-MSC与肝分化的hUC-MSC之间至少改变了2倍。在这些microRNA中,有25个过表达;这种过度表达是逐渐发生或急剧增加,并保持在较高水平。共有36个microRNA表达不足,其表达模式与过度表达的microRNA相似。还通过定量逆转录聚合酶链反应证实了改变的表达的microRNA的表达。我们还发现,参与肝细胞分化的microRNA在肝细胞或肝细胞癌细胞中并未富集,并且可能靶向肝富集的转录因子和基因。阐明在hUC-MSC肝分化过程中的microRNA谱为澄清microRNA在hUC-MSC肝分化中的作用以及将hUC-MSC转化为肝细胞的特异性microRNA选择提供了基础。

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