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Expression data from iPS and human ES cells

ABSTRACT: Assays to assess the quality of the reprogramming of human Induced pluripotent stem cells are needed. We have previously shown that hESC can be differentiated into embryonic and fetal type of red blood cells that sequentially express three types of hemoglobins recapitulating early human erythropoiesis. We report here that we have produced iPS from three somatic cell types: adult skin fibroblasts as well as embryonic and fetal mesenchymal stem cells. We show that some of these iPS are fully reprogrammed into a pluripotent state that is undistinguishable from that of hESCs based and low and high-throughput expression analysis and detailed expression of globin expression patterns suggesting that reprogramming with the four original Yamanaka pluripotency factors leads to complete erasure of all functionally important epigenetic marks associated with hematopoietic differentiation regardless of the age or the tissue type of the donor cells. We also report that reprogramming can also lead to abnormal iPS that are undistinguishable from hES cells by morphology, and by the expression of their endogenous genes but that are grossly abnormal in their differentiation potential. The most likely cause of abnormal reprogramming is failure to silence the virally transduced reprogramming factors. The ability to produce large number of erythroid cells with embryonic and fetal-like characteristics is likely to have many translational applications

ORGANISM(S): Homo sapiens

PROVIDER: GSE26946 | GEO | 2011/01/29

SECONDARY ACCESSION(S): PRJNA136947

REPOSITORIES: GEO

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Expression data from iPS and human ES cells

Project description:Assays to assess the quality of the reprogramming of human Induced pluripotent stem cells are needed. We have previously shown that hESC can be differentiated into embryonic and fetal type of red blood cells that sequentially express three types of hemoglobins recapitulating early human erythropoiesis. We report here that we have produced iPS from three somatic cell types: adult skin fibroblasts as well as embryonic and fetal mesenchymal stem cells. We show that some of these iPS are fully reprogrammed into a pluripotent state that is undistinguishable from that of hESCs based and low and high-throughput expression analysis and detailed expression of globin expression patterns suggesting that reprogramming with the four original Yamanaka pluripotency factors leads to complete erasure of all functionally important epigenetic marks associated with hematopoietic differentiation regardless of the age or the tissue type of the donor cells. We also report that reprogramming can also lead to abnormal iPS that are undistinguishable from hES cells by morphology, and by the expression of their endogenous genes but that are grossly abnormal in their differentiation potential. The most likely cause of abnormal reprogramming is failure to silence the virally transduced reprogramming factors. The ability to produce large number of erythroid cells with embryonic and fetal-like characteristics is likely to have many translational applications the gene expression in different source of iPS and standard ES cells H1

2011-01-29 | E-GEOD-26946 | biostudies-arrayexpress

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Comparisons between fully and partially reprogrammed iPS cells induced by pMX-Klf4, pMX-Oct4 and pMX-Sox2 retroviruses

Project description:Induced pluripotent stem (iPS) cell reprogramming is a gradual epigenetic process that reactivates the pluripotent transcriptional network by erasing and establishing heterochromatin marks. Here, we characterize the physical structure of heterochromatin domains in full and partial mouse iPS cells by correlative Electron Spectroscopic Imaging (ESI). In somatic and partial iPS cells, constitutive heterochromatin marked by H3K9me3 is highly compartmentalized into chromocenter structures of densely packed 10 nm chromatin fibers. In contrast, chromocenter boundaries are poorly defined in pluripotent ES and full iPS cells, and are characterized by unusually dispersed 10 nm heterochromatin fibers in high Nanog-expressing cells, including pluripotent cells of the mouse blastocyst prior to differentiation. This heterochromatin reorganization accompanies retroviral silencing during conversion of partial iPS cells by Mek/Gsk3 2i inhibitor treatment. Thus, constitutive heterochromatin reorganization serves as a novel biomarker with retroviral silencing for identifying iPS cells in the very late stages of reprogramming. We compared the expression profiles of partially and fully reprogrammed iPS cell lines derived from CD1 mouse embryonic fibroblasts (MEFSs) by retroviral reprogramming (pMX-Oct4, pMX-Klf4 and pMX-Sox2). to the differentiated MEFS and the J1 embryonic stem cell line. We also studied the effect of a 2i cocktail treatment in partially reprogrammed iPS cells.

2011-09-21 | E-GEOD-21595 | biostudies-arrayexpress

Expression data from mouse ES and iPS cells

Project description:Induced pluripotent stem (iPS) cells were produced from reprogramming of somatic cells, and they are shown to possess pluripotent properties similar to embryonic stem (ES) cells. Here we used microarrays to detail the global expression pattern among the ES cells and iPS cells, as well as the original mouse embryo fibroblast (MEF), to identify important players involved in the reprogramming process.

2010-05-13 | GSE21515 | GEO

Expression data from mouse ES and iPS cells

2009-08-03 | GSE16925 | GEO

Project description:The ability to induce pluripotent stem cells from committed, somatic human cells provides tremendous potential for regenerative medicine. However, there is a defined neoplastic potential inherent to such reprogramming that must be understood and may provide a model for understanding key events in tumorigenesis. Using genome-wide assays, we identify cancer-related epigenetic abnormalities that arise early during reprogramming and persist in induced pluripotent stem cell (iPS) clones. These include hundreds of abnormal gene silencing events, patterns of aberrant responses to epigenetic-modifying drugs resembling those for cancer cells, and presence in iPS and partially reprogrammed cells of cancer-specific gene promoter DNA methylation alterations. Our findings suggest that by studying the process of induced reprogramming, we may gain significant insight into the origins of epigenetic gene silencing associated with human tumorigenesis, and add to means of assessing iPS for safety. Direct expression comparison of iPS lines, cultured stem cell lines and normal differentiated cells. Re-expression experiments with 5-aza-2′-deoxycytidine (AZA) and trichostatin A (TSA) to identify hypermethylated genes.

2011-06-16 | E-GEOD-29870 | biostudies-arrayexpress

Reprogramming human somatic cells to a pluripotent state

Project description:Here we compare the global gene expression of induced pluripotent stem cell (iPS) cells with those of normal human embryonic stem cells and parental mesenchymal cells. Keywords: reprogramming, iPS We used 1 chip/sample (3 iPS clones for each mesenchymal clone - 12 iPS clones; 4 mesenchymal clones and 5 normal human ES cell lines).

2010-05-27 | E-GEOD-9071 | biostudies-arrayexpress

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Uniform, integration-free iPS cells with a safeguard system using human artificial chromosome vectors

Project description:Human artificial chromosomes (HACs) have unique characteristics as gene-delivery vector, e.g., episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) to induced pluripotent stem (iPS) cells. Two HAC vectors (iHAC1 and iHAC2) were constructed. Both carried four reprogramming factors, and iHAC2 also encoded a p53-knockdown construct (see Kazuki et al 2011). The iHAC1 partially reprogrammed MEFs, and the iHAC2 efficiently reprogrammed MEFs. Global gene expression showed that the iHACs, unlike other vectors, generated relatively uniform iPS cells. We established iHAC-free iPS cells by isolating cells that spontaneously lost the iHAC2. Analyses of pluripotent markers, teratomas, and chimeras confirmed that these iHAC-free iPS cells were pluripotent. Moreover, iHAC-free iPS cells with a re-introduced HAC encoding Herpes Simplex Virus Thymidine Kinase were eliminated by Ganciclovir treatment; therefore, the HAC safeguard system functioned in iPS cells. Thus, the HAC vector could generate uniform, integration-free iPS cells with a safeguard system. Total RNA from mouse embryonic fibroblasts, two control ES cells (TT2 and B6ES), retrovirus vector-derived iPS cells (20D17), and twelve HAC-derived iPS clones were hybridized to the Agilent Whole Mouse Genome microarrays.

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