title: Epigenetic regulation of human adipose-derived stem cells differentiation.
abstract: Adult stem cells have more restricted differentiation potential than embryonic stem cells (ESCs), but upon appropriate stimulation can differentiate into cells of different germ layers. Epigenetic factors, including DNA modifications, take a significant part in regulation of pluripotency and differentiation of ESCs. Less is known about the epigenetic regulation of these processes in adult stem cells. Gene expression profile and location of DNA modifications in adipose-derived stem cells (ADSCs) and their osteogenically differentiated lineages were analyzed using Agilent microarrays. Methylation-specific PCR and restriction-based quantitative PCR were applied for 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) detection in selected loci. The level of DNA modifications in the POU5F1 locus was quantified with deep sequencing. Expression levels of selected genes were assayed by real-time PCR. ADSCs differentiation into osteogenic lineages involved marked changes in both 5mC and 5hmC profiles, but 5hmC changes were more abundant. 5mC losses and 5hmC gains were the main events observed during ADSCs differentiation, and were accompanied by increased expression of TET1 (P = 0.009). In ADSCs, POU5F1 was better expressed than NANOG or SOX2 (P ≤ 0.001). Both 5mC and 5hmC marks were present in the POU5F1 locus, but only hydroxymethylation of specific cytosine showed significant effect on the gene expression. In summary, the data of our study suggest significant involvement of changes in 5hmC profile during the differentiation of human adult stem cells.
authors: Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University, Ciurlionio 21, 03101, Vilnius, Lithuania.
authors: Kristina
authors: K
authors: Daniunaite
authors: Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University, Ciurlionio 21, 03101, Vilnius, Lithuania.
authors: Inga
authors: I
authors: Serenaite
authors: Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University, Ciurlionio 21, 03101, Vilnius, Lithuania.
authors: Roberta
authors: R
authors: Misgirdaite
authors: Institute of Mathematics and Informatics, Vilnius University, Akademijos 4, 08663, Vilnius, Lithuania.
authors: Juozas
authors: J
authors: Gordevicius
authors: State Research Institute Center of Innovative Medicine, Zygimantu 9, 01120, Vilnius, Lithuania.
authors: Ausra
authors: A
authors: Unguryte
authors: EFS Pyreenees-Mediterranee, 31027, Toulouse, France.|UMR5273 CNRS, EFS-INSERM U1031, Stromalab, 31432, Toulouse, France.
authors: Sandrine
authors: S
authors: Fleury-Cappellesso
authors: State Research Institute Center of Innovative Medicine, Zygimantu 9, 01120, Vilnius, Lithuania.
authors: Eiva
authors: E
authors: Bernotiene
authors: Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University, Ciurlionio 21, 03101, Vilnius, Lithuania. sonata.jarmalaite@gf.vu.lt.
authors: Sonata
authors: S
authors: Jarmalaite
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism / Q000031:analogs & derivatives; D000273:Adipose Tissue / Q000166:cytology*; D053687:Adult Stem Cells / Q000378:metabolism / Q000502:physiology*; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D019070:Cell Lineage; D003596:Cytosine / Q000031:analogs & derivatives / Q000378:metabolism; D044127:Epigenesis, Genetic; D020869:Gene Expression Profiling / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000378:metabolism / Q000502:physiology*; D010012:Osteogenesis / Q000235:genetics*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D011518:Proto-Oncogene Proteins; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D006899:Mixed Function Oxygenases; C462539:TET1 protein, human
keywords: 5-hydroxymethylcytosine; Adipose-derived stem cells; POU5F1