title: Higher methylation in genomic DNA indicates incomplete reprogramming in induced pluripotent stem cells.
abstract: Pluripotent stem cells can be created successfully through the inner cell mass (ICM), nuclear transfer, and defined-factor induction. Unfortunately, the epigenetic characteristics of the cells produced are poorly understood. In this article, we compared expression levels of enzymes involved in epigenetic modifications across six pluripotent stem cell lines. Six of the 11 genes evaluated here (Dnmt3a, Dnmt3b, Tet1, Ezh2, Mll1, and Lsd1) showed abnormally low levels of expression in the two germ-line chimeric induced pluripotent stem cell (iPSC) lines. We also conducted locus-specific analysis of DNA methylation at 9 loci. Although iPSCs did express Oct4, the Oct4 promoter region was shown to have a higher level of DNA methylation. The Xist and Line-1 repeating sequences differed relatively little in methylation level across the cell lines, but Peg3, Peg10, and H19 exhibited high degrees of variation in the pattern of DNA methylation. Meg3 in the Dlk1-Dio3 imprinting cluster was incompletely methylated in embryonic stem cells (ESCs) and nuclear transfer (nt) ESCs. However, in germ-line chimeric iPSCs, Meg3 was almost entirely methylated. ESC and ntESC lines showed twice as much Meg3 expression than in the iPSC lines. The genomic 5mC contents detected by reverse-phase high-performance liquid chromatography (HPLC) indicated that, despite their germ-line chimeric abilities, iPSCs remained incompletely reprogrammed, even though no direct evidence is shown here.
authors: School of Life Sciences, Henan University, Kaifeng, PR China.
authors: Wenping
authors: W
authors: Zhou
authors: Kun
authors: K
authors: Wang
authors: Weimin
authors: W
authors: Ruan
authors: Zhang
authors: Z
authors: Bo
authors: Lifang
authors: L
authors: Liu
authors: Zhongyang
authors: Z
authors: Cao
authors: Lianqin
authors: L
authors: Chai
authors: GengSheng
authors: G
authors: Cao
mesh_terms: D000818:Animals; D054337:Cell Dedifferentiation; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D005786:Gene Expression Regulation / Q000235:genetics; D056426:Genetic Loci; D018392:Genomic Imprinting; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D005810:Multigene Family / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
chemical_list: D014157:Transcription Factors; D004247:DNA
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