|
| Status |
Public on Sep 24, 2012 |
| Title |
MSC donor11 P14 hTERT |
| Sample type |
genomic |
| |
|
| Source name |
mesenchymal stromal cells (MSC)
|
| Organism |
Homo sapiens |
| Characteristics |
donor: donor11
cell type: mesenchymal stromal cells (MSC)
passage: P14
|
| Treatment protocol |
MSC were isolated from tibia plateau of patients undergoing orthopaedic surgery. All samples were taken after written consent using guidelines approved by the Ethic Committee on the Use of Human Subjects at the University of Aachen (Permit number: EK128/09). MSC were culture-expanded in standard medium consisting of DMEM (PAA; 1g/L glucose) supplemented with glutamine (PAA), penicillin/streptomycin (PAA) and 10% human platelet lysate (hPL).
|
| Growth protocol |
The hTERT sequence was cloned into the retroviral expression vector pMIG (pMSCV-IRES-GFP) (Addgene plasmid 9044, Addgene, Cambridge, MA, USA) via EcoRI. For control we used the corresponding empty pMIG vector (IRES-GFP). Transfected MSC were culture expanded in parallel until entering the senescent state.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Genomic DNA extraction
|
| Label |
Cy5 and Cy3
|
| Label protocol |
standard Illumina labeling protocol
|
| |
|
| Hybridization protocol |
Bisulfite converted DNA was amplified, fragmented and hybridized to Illumina Infinium HumanMethylation450 Beadchip using standard Infinium HD Methylation Assay protocol.
|
| Scan protocol |
Standard Infinium HD Methylation Assay protocol
|
| Data processing |
Initial analysis was performed by the Genomestudio 2010.3 (Modul M Version 1.8.5). Data were normalized with internal controls according to Illumina´s standard procedures. Methylation level at each locus was calculated with the GenomeStudio Methylation module as beta-value (ranging from 0 to 1). The number of beads per feature varies between chips and beta-values were calculated as average of at least three technical replica.
|
| |
|
| Submission date |
Apr 05, 2012 |
| Last update date |
Sep 24, 2012 |
| Contact name |
Wolfgang Wagner |
| E-mail(s) |
wwagner@ukaachen.de
|
| Phone |
+49 241 8088611
|
| Organization name |
RWTH Aachen University
|
| Department |
Helmholtz Institute for Biomedical Engineering
|
| Lab |
Stem Cell Biology and Cellular Engineering
|
| Street address |
Pauwelsstrasse 20
|
| City |
Aachen |
| ZIP/Postal code |
52074 |
| Country |
Germany |
| |
|
| Platform ID |
GPL13534 |
| Series (2) |
| GSE37066 |
Pluripotent Stem Cells Escape From Senescence-Associated DNA Methylation Changes [Illumina] |
| GSE37067 |
Pluripotent Stem Cells Escape From Senescence-Associated DNA Methylation Changes |
|
