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Methylation Pattern of Human Dermal Fibroblasts

ABSTRACT: In this study, we have analyzed DNA methylation changes upon aging of human dermal fibroblasts by using the HumanMethylation27 BeadChip assessing 27,578 unique CpG sites. Cells were isolated from skin samples donated by young (6-23 years) and elderly (60-73 years) patients undergoing surgical interventions. Strikingly, global DNA-methylation profiles of fibroblasts from the same anatomical site clustered closely together indicating that fibroblasts maintain positional memory even after in vitro culture.

ORGANISM(S): Homo sapiens

PROVIDER: GSE22595 | GEO | 2012/02/14

SECONDARY ACCESSION(S): PRJNA128371

REPOSITORIES: GEO

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Methylation Pattern of Human Dermal Fibroblasts

Project description:In this study, we have analyzed DNA methylation changes upon aging of human dermal fibroblasts by using the HumanMethylation27 BeadChip assessing 27,578 unique CpG sites. Cells were isolated from skin samples donated by young (6-23 years) and elderly (60-73 years) patients undergoing surgical interventions. Strikingly, global DNA-methylation profiles of fibroblasts from the same anatomical site clustered closely together indicating that fibroblasts maintain positional memory even after in vitro culture. Skin samples from younger or elderly donors were treated with dispase (Roche Diagnostics, Mannheim, Germany) for 12 hours to separate the dermis from the epidermis. The dermis was digested with 0,2% collagenase and 1,5% BSA in collagenase buffer (100mM HEPES, 120mM NaCl, 50mM KCl, 1mM CaCl2, 5mM Glucose) for 45 minutes. Dermal remnants were removed by filtering the digest through a 100Âµm nylon strainer (Falcon, Becton Dickinson [BD], San Jose, USA). The cells were subsequently washed and expanded in standard medium consisting of DMEM (PAA; 1g/L glucose) supplemented with glutamine (PAA), penicillin/sptrepamycin (PAA) and 10% fetal calf serum (Biochrom, Berlin, Germany) in a humidified atmosphere at 5% CO2. Cells were always replated when grown to 80% confluency. For methylation profiles upon aging we have isolated DNA from the samples of passage 3 of younger and elderly donors. For methylation profiles upon long-term culture we have isolated DNA from the samples of early passage (P3) and late passage (P21)

2012-02-14 | E-GEOD-22595 | biostudies-arrayexpress

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Project description:Genome wide DNA methylation profiling of tumor samples with lung adenocarcinoma patients. The Illumina HumanMethylation27 BeadChip array was used to obtain DNA methylation profiles across approximately 27,578 CpGs in lung cancer samples. Samples included tumor samples with lung adenocarcinoma.

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Epigenetic analysis of gastric cancer

Project description:Genome-wide DNA methylation profiling of gastric tumors and matched gastric non-malignant samples. The Illumina HumanMethylation27 BeadChip was used to obtain DNA methylation profiles across 27,578 CpGs in 203 gastric tumors and 94 matched non-malignant gastric samples. Bisulphite-converted DNA from the 203 gastric tumors and 94 matched gastric non-malignant samples was hybridized to the Illumina HumanMethylation27 BeadChip.

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Methylation Pattern of Human Mesenchymal Stromal Cells

Project description:In this study, we have analyzed DNA methylation changes upon long-term culture and aging of MSC by using the HumanMethylation27 BeadChip assessing 27,578 unique CpG sites. Cells were taken from bone marrow aspirates from iliac crest (BM) of healthy donors or from the caput femoris (HIP) of elderly patients that received femoral head prosthesis.Overall, the methylation pattern was maintained throughout both long-term culture and aging but highly significant differences were observed at specific CpG sites. Notably, methylation changes in MSC were related in long-term culture and aging in vivo. Mesenchymal stromal cells (MSC) were isolated from human bone marrow as described before (Wagner et al., 2006, Exp Hematol, 34, 536-548; Wagner et al.,2008, PLoS One 21;3(5):e2213.; Wagner et al., PLoS One. 2009 Jun 9;4(6):e5846.) Cells were always replated when grown to confluency. A sample for DNA isolation was taken at every passage until the cells finally became senescent. For methylation profiles upon long-term culture we used the samples of early passage (P2) and senescent passage (PX). For methylation profiles upon aging we have only used the samples of passage 2 of younger and elderly donors. Gene expression data of these samples have been previously deposited unter GSE9593 and GSE12274.

2010-05-14 | E-GEOD-17448 | biostudies-arrayexpress

Genome wide DNA Methylation Study in Cornelia de Lange Syndrome

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2010-07-07 | E-GEOD-18458 | biostudies-arrayexpress

A genome-wide methylation study on essential hypertension in young African American Males

Project description:We performed a genome wide methylation scan using the Illumina HumanMethylation27 BeadChip on DNAs extracted from the leukocytes of 8 hypertensive cases and 8 normotensive age-matched controls. All subjects were African American males aged 14-23 years.

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Project description:We performed a genome wide methylation scan using the Illumina HumanMethylation27 BeadChip on DNAs extracted from the leukocytes of 7 obese (BMI≥99th percentile for age and sex) and 7 lean (BMI≤10th percentile for age and sex) African American males aged 14-18 years.

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Project description:Genome wide DNA methylation profiling of muscle cells extracted from healthy donors at different ages. The Illumina Infinium Human Methylation 450 BeadChip was used to obtain DNA methylation profiles across > 485,000 methylation sites. The muscle cells were extracted from young adults (18-40 years old) and elderly (>70 years old)(n=3 per group, 2 groups : Young and aged adult). Bisulphite converted DNA from the 6 samples were hybridized to the Illumina Infinium Human Methylation 450 BeadChip

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