Dataset Information


Human induced pluripotent stem (iPS) cells from neonatal skin derived cells

ABSTRACT: Induction of germline-competent pluripotent stem cells from mouse fibroblasts has been achieved by the ectopic expression of four genes (Oct3/4, Sox2, c-Myc and Klf4). If this method can be applied to humans for the generation of personalized human pluripotent stem cells, it would greatly facilitate the therapeutic application of stem cells by avoiding the problem of immune rejection by the recipient associated with allograft transplants. Here we show that the ectopic expression of the same four genes in human neonatal skin derived cells is sufficient to induce pluripotent stem cells indistinguishable from human embryonic stem cells in morphology, gene expression, DNA methylation, teratoma formation and long term self-renewal ability. Extensive analysis of colonies generated by ectopic expression of these four genes indicates the presence of considerable heterogeneity in the induced colonies. These results provide a new finding to generate human induced pluripotent stem cells from postnatal somatic tissues. Keywords: Cell type comparison Overall design: The microarray study was carried out using Affymetrix Human Genome U133 Plus 2.0 gene expression arrays or Agilent Whole Human Genome Oligo microarrays. Total RNA was extracted from cells with RNeasy (Qiagen). For Affymetrix array, biotin-labelled cRNA was reverse transcribed from 1 µg of total RNA according to Affymetrix technical protocols. Fifteen micrograms of cRNA was fragmented and hybridized to Affymetrix U133 plus 2 GeneChip arrays at 45C for 16 h and then washed and stained using Affimetrix Fluidics. The arrays were scanned using an Affimetrix GCS3000 scanner, and the images obtained were processed to data using the GCOS software. For Agilent array (G4112A), Total RNA was labeled with Cy3. Samples were hybridized with Agilent Gene Expression Hybridization kit according to the one color protocol. Arrays were scanned with a Agilent Microarray Scanner System and processed to data with Feature Extraction Software (v.9.5.3). Data from this experiment and the GEO database were analysed with the GeneSpring 7.3.1. software.


INSTRUMENT(S): [HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array

ORGANISM(S): Homo sapiens  

SUBMITTER: Tetsuya Ishikawa  

PROVIDER: GSE9709 | GEO | 2008-03-16



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