MSCs were isolated from bone marrow (tibia plateau) after written consent using guidelines approved by the Ethic Committee of the Use of Human Subjects at the University of Aachen (permit number: EK128/09). MSCs from three donors were infected with pMXs based retroviruses (Addgene, Cambridge, MA, USA) carrying the OCT3/4, SOX2, KLF4, and c-MYC genes. Established iPSCs were maintained on MEFs in DMEM/F12 medium supplemented with Glutamax, 20% knockout serum replacer, 1% nonessential amino acids, 1x penicillin/streptomycin, 1x L-glutamine, 0.1 mM β-Mercaptoethanol, and 50 ng/ml basic fibroblast growth factor (Peprotech, Hamburg, Germany). To exclude contamination of feeder layer cells, iPSCs were adjusted to a feeder-free system on matrigel (BD Biosciences, San Jose, CA, USA) in mTeSR™1 medium (Stemcell Technologies, Vancouver, BC, Canada) for at least three passages. Cells were passaged every 5-6 days enzymatically with dispase (1 g/ml, Stemcell Technologies). For re-differentiation of iPSC towards iPS-MSCs medium was simply exchanged for MSC standard medium supplemented with 10% human platelet lysate (hPL) for 7 days, and cells were then further passaged in culture wells coated with 0.1% gelatin (Sigma-Aldrich, St. Louis, CA, USA).
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated with the NucleoSpin® miRNA Isolation Kit (Macherey-Nagel, Düren, Germany).
Label
biotin
Label protocol
Affymetrix protocol
Hybridization protocol
Affymetrix protocol
Scan protocol
Arrays were scanned using Affymetrix GeneChip Scanner 3000.
Data processing
CEL files were processed using the Affymetrix Power Tools (APT) Software Package. Normalization was done employing RMA (Irizarry et al., Biostatistics 4, 249-264, 2003).
Tissue- and Aging-specific DNA-Methylation Patterns are erased in Mesenchymal Stromal Cells derived from Induced Pluripotent Stem Cells. [Expression profiling]
