Viral and transgene-free human induced pluripotent stem (iPS) cells using non-integrating Sendai virus (SeV) vectors.
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ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Noemi Fusaki
PROVIDER: E-GEOD-24240 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Reprogramming of somatic cells provides potential for the generation of specific cell types, which could be a key step in the study and treatment of human diseases. In vitro reprogramming of somatic cells into a pluripotent embryonic stem (ES) cellM-bM-^@M-^Slike state has been reported by retroviral transduction of murine fibroblasts using four embryonic transcription factors or through cell fusion of somatic and pluripotent stem cells. The generation of reprogrammed pluripotent cells using a somatic cell donor source such as bone marrow (BM) or peripheral blood is of particular therapeutic interest because of the relative ease of harvesting these cell types. Here we show that mouse adult BM mononuclear cellsM-oM-<M-^HBM MNCsM-oM-<M-^Iare competent as donor cells and can be reprogrammed into pluripotent ES cell-like cells. We isolated BM MNCs and embryonic fibroblasts (MEFs) from Oct4-EGFP transgenic mice, fused them with ES cells and infected them with retroviruses expressing Oct4, Sox2, Klf4, and c-Myc. Fused BM cells formed more ES-like colonies than did MEFs. Infected BM cells gave rise to iPS cells, although transduction efficiencies were not high. It was more efficient to pick up iPS colonies as compared with MEFs. BM-derived iPS (BM iPS) cells expressed embryonic stem cell markers, formed teratomas, and contributed to chimera mice with germline development. Clonal analysis revealed that BM iPS clones had diversity, although some clones were found to be genetically identical with different phenotypes. Here we demonstrate, for the first time, the induction of pluripotent cells directly from hematopoietic tissue. Gene expression profiling was performed in mouse BMMNCs, ES and BMMNC derived iPS cell lines.
2009-07-02 | E-GEOD-15775 | biostudies-arrayexpress
Project description:Human induced pluripotent stem cells (iPS cells) resemble embryonic stem cells and can differentiate into cell derivatives of all three germ layers. However, frequently the differentiation efficiency of iPS cells into some lineages is rather poor. Here, we found that fusion of iPS cells with human hematopoietic stem cells (HSC) enhances iPS cell differentiation. Such iPS hybrids showed a prominent differentiation bias towards hematopoietic lineages but also towards other mesendodermal lineages. Additionally, during differentiation of iPS hybrids expression of early mesendodermal markers - Brachyury (T), MIX1 Homeobox-Like Protein 1 (MIXL1) and Goosecoid (GSC) - appeared with faster kinetics than in parental iPS cells. Following iPS hybrid differentiation there was a prominent induction of NODAL and inhibition of NODAL signaling blunted mesendodermal differentiation. This indicates that NODAL signaling is critically involved in mesendodermal bias of iPS hybrid differentiation. In summary, we demonstrate that iPS cell fusion with HSC prominently enhances iPS differentiation. 11 samples were hybridized GeneChip Human Gene 1.0 ST Arrays (Affymetrix)
2014-08-06 | E-GEOD-58457 | biostudies-arrayexpress
Project description:Induced pluripotent stem (IPS) cells have attracted enormous attention due to their vast potential in regenerative medicine, pharmaceutical screening and basic research. The majority of prior established rat IPS cells were generated from somatic cells by retroviral and lentiviral transduction with expression of Oct4, Sox2, Klf4 and c-Myc and using chemical inhibitors of key differentiation pathways. A major difficulty in the application of this technology is the efficient delivery of reprogramming factors and the long-term maintenance of properties of stem cells. Here, we employed the PiggyBac (PB) transposon carrying four 2A peptide-linked reprogramming factors for generating rat IPS cells. These stable rat IPS cells are similar to embryonic stem (ES) cells in morphology, proliferation, teratoma formation, expression characteristic pluripotency markers, developmental potential, and germline transmission. Transcriptional profiling of the IPS cells revealed both pathways in common with ES cells from rat and unique signaling pathway to our cells, including Wnt, TGF and Notch. The cell lines and information obtained in this study will accelerate our understanding of the molecular regulation underlying germline pluripotency and pave the way for exploration of cell-based therapies using the rat. To compare the gene expression profiling between rat IPS cells and ES cells to show if the rat IPS cells had been reprogrammed into pluripotent status like rat ES cells at the gene expression level.
2012-02-16 | E-GEOD-35841 | biostudies-arrayexpress
Project description:Microarray experiment 2: comparison of mRNA expression levels between iPS clones #1, 4, 5 and ES cells
2013-03-22 | E-TABM-1218 | biostudies-arrayexpress
Project description:Owing to the risk of insertional mutagenesis, viral transduction has been increasingly replaced by nonviral methods to generate induced pluripotent stem (iPS) cells. We report the use of âminicircleâ DNA, a vector type that is free of bacterial DNA and capable of high expression in cells. Here we use a single minicircle vector to generate transgene-free iPSCs from adult human adipose stem cells. (Note: Our Nature Methods publication included analysis of array data from GSM378832 (Foreskin), GSM378833-GSM378838 (JT-iPSC), and GSM378817-GSM378820 (H1, H7, H9, H13, H14) in conjunction with this series). Total RNA from human adipose stem cells (hASC, n = 3 replicate samples), hASC-derived iPS cells using lentiviral factors (lenti-iPSC, n = 3 replicate samples), and minicircle-derived human iPS cells (mc-iPSC, n = 3 subclones from adipose tissue of three individual patients) was hybridized to nine Affymetrix GeneChip Human Genome U133 Plus 2.0 Arrays.
2010-02-21 | E-GEOD-20033 | biostudies-arrayexpress
Project description:We were able to achieve an initial stable intermediate phase by the transduction of Oct4, Klf4, and c-Myc. Furthermore, over-expression of Sox2 in these intermediate stage cells leads to final iPS cell phase. After examining the gene expression profiles from the initial to final iPS cell phases, we have identified Sox2 downstream genes important for iPS cell induction. 4 groups of cells are analyzed. Each group contains cells from 4 indepent dishes. The four groups are: ES cells, iPS cells, intermediate stage cells (OKM cells) and MEFs
2011-12-29 | E-GEOD-28197 | biostudies-arrayexpress
Project description:Whole genome sequencing was performed on several murine iPS cell clones (and their parental cells) from each of three independent reprogramming experiments. Hundreds of single nucleotide variants (SNVs) were detected in each clone, with an average of 11 in coding regions. Affymetrix Mouse Exon 1.0ST arrays were used to compare expression patterns in MPSVII iPS lines, and embryo-derived MPSVII ES cells. Unsupervised hierarchal clustering analysis showed that the iPS clones and ES cell lines clustered randomly, suggesting that their global patterns of gene expression are highly similar. Taken together, our data suggest that most of the genetic variation in iPS cell clones is not caused by reprogramming, but is rather a consequence of cloning individual cells, “capturing” random mutations that preexisted in the single cells that were reprogrammed. These mutations can sometimes contribute to reprogramming “fitness”, thus providing a selective advantage for rare cells when they overexpress reprogramming factors. Mouse embryonic fibroblasts (MEFs) derived from a murine disease model (Mucopolysaccaridosis type VII- MPSVII) were used. Affymetrix Mouse Exon 1.0ST arrays were used to compare expression patterns in MPSVII iPS lines, and embryo-derived MPSVII ES cells. Expression patterns in four separate iPS clones were compared to MPSVII ES cells. Control hybridization was performed with B6 Blu ES cells and MEFs.
2012-03-15 | E-GEOD-36017 | biostudies-arrayexpress
Project description:To reprogram mouse embryonic fibroblasts (MEFs) to induced Pluripotent Stem Cells (iPSCs), we constructed the PiggyBac (PB) transposon carrying the four Yamanaka factor cDNAs controlled by a CAG promoter (PB-CAG-OCKS, Oct4, cMyc, Klf4 and Sox2). As the baseline reprogramming control, we transfected the PB-CAG-OCKS transposon into Oct4-reporter MEFs and plated the cells on STO feeder cells (4F-iPS). To examine the effects of miR-25 on reprogramming, in addition to the Yamanaka factors, we co-transfected the PB-CAG-OCKS plasmid with the PB-CAG-miR-25 plasmid and selected for puromycin resistance (2.0 mg/ml) (25-iPS). We then performed genome-wide gene expression microarray analysis on the iPS cells generated and compared the expression profiles to those of Oct4-reporter MEFs and wildtype ES cells.
2011-10-10 | E-MTAB-409 | biostudies-arrayexpress
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. Experiment Overall Design: Mouse ES cell cultures, as well as selected iPS cell lines and the original MEF cells they were derived from, were used for RNA extraction and hybridization on Affymetrix microarrays. Three biological replicates for each sample were processed. GeneChips were processed and data were analyzed as previously described (Zeng et al., Dev Biol 20004).
2009-08-08 | E-GEOD-16925 | biostudies-arrayexpress
Project description:Induced pluripotent stem (iPS) cells have been derived from various somatic cell populations through ectopic expression of defined factors. It remains unclear whether iPS cells generated from different cell types are molecularly and functionally similar. Here, we show that iPS cells obtained from fibroblasts, hematopoietic and myogenic cells exhibit distinct transcriptional and epigenetic patterns. Moreover, we demonstrate that cellular origin influences the in vitro differentiation potentials of iPS cells into embryoid bodies and different hematopoietic cells. Our results suggest that low-passage iPS cells retain a transient epigenetic memory of their somatic cells of origin, which manifests as differential gene expression and altered differentiation capacity. These observations might affect ongoing attempts to use iPS cells for disease modeling and also could be exploited for potential therapeutic applications to enhance differentiation into desired cell lineages. This series consists of triplicated mRNA expression microarray data (Affymetrix mouse gene ST 1.0) for an early passage (passage 4) of mouse iPS cells derived from bone marrow granulocytes, splenic B cells, tail tip fibroblasts, and skeletel muscle precursor cells. iPS cells were generated by infecting somatic mouse cells with lentiviruses expressing Oct4, Sox2, Klf4, and cMyc. Total RNA was isolated from iPS cells at different passages.
2010-07-19 | E-GEOD-22908 | biostudies-arrayexpress