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Human induced pluripotent stem cells free of exogenous DNA are derived with episomal vectors (fig 4.a)

ABSTRACT: Human induced pluripotent stem (iPS) cells have previously been derived from somatic cells using viral vectors that integrate transgenes into the genome. Genomic integration, however, can allow persistent leaky expression of the transgenes and can create insertional mutations, thus limiting the utility of these cells for both research and clinical applications. Here, we describe the derivation of human iPS cells free of vector and transgene sequences using non-integrating oriP/EBNA1-based episomal vectors. The resulting iPS cells are similar to human embryonic stem (ES) cells in both proliferative and developmental potential. These results demonstrate that reprogramming of human somatic cells does not require genomic integration or the continued presence of exogenous reprogramming factors, and removes one important obstacle to the clinical applications of these cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE15176 | GEO | 2009/03/27

SECONDARY ACCESSION(S): PRJNA123129

REPOSITORIES: GEO

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Human induced pluripotent stem cells free of exogenous DNA are derived with episomal vectors (fig 1.c)

2009-03-27 | GSE15175 | GEO

Transcription profiling of human induced pluripotent stem cells free of exogenous DNA are derived with episomal vectors (fig 1.c)

2009-04-04 | E-GEOD-15175 | biostudies-arrayexpress

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Human induced pluripotent stem cells free of exogenous DNA are derived with episomal vectors

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Transcription profiling of human induced pluripotent stem cells free of exogenous DNA are derived with episomal vectors (super series)

Project description:Human induced pluripotent stem (iPS) cells have previously been derived from somatic cells using viral vectors that integrate transgenes into the genome. Genomic integration, however, can allow persistent leaky expression of the transgenes and can create insertional mutations, thus limiting the utility of these cells for both research and clinical applications. Here, we describe the derivation of human iPS cells free of vector and transgene sequences using non-integrating oriP/EBNA1-based episomal vectors. The resulting iPS cells are similar to human embryonic stem (ES) cells in both proliferative and developmental potential. These results demonstrate that reprogramming of human somatic cells does not require genomic integration or the continued presence of exogenous reprogramming factors, and removes one important obstacle to the clinical applications of these cells. This SuperSeries is composed of the following subset Series:; GSE15175: Human induced pluripotent stem cells free of exogenous DNA are derived with episomal vectors (fig 1.c); GSE15176: Human induced pluripotent stem cells free of exogenous DNA are derived with episomal vectors (fig 4.a) Experiment Overall Design: Total 21 samples were analyzed to confirm the similarity of human iPS cells derived with episomal vectors with human ES cells, and a dissimilarity with fibroblasts. Experiment Overall Design: Refer to individual Series

2009-04-04 | E-GEOD-15148 | biostudies-arrayexpress

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.

2011-10-16 | GSE29441 | GEO

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

2011-10-15 | E-GEOD-29441 | biostudies-arrayexpress

Generation of Integration-free Induced Neural Stem Cells from Mouse Fibroblasts

Project description:Recent studies demonstrated that fibroblasts could be converted into induced neural stem cells (iNSCs). However, the insertional mutation caused by random integration of viral vectors has been a major limitation of iNSCs for the future clinical translation. Here we show that non-viral transfection of episomal vectors encoding Brn4/Pou3f4, Sox2, Klf4, and c-Myc sufficiently generates iNSCs. The episomal vector mediated iNSCs closely resemble brain-derived NSCs as well as iNSCs generated by retrovirus in morphology, gene expression profile, epigenetic status, self-renewal capacity and both in vitro and in vivo differentiation capacity. The novel conversion protocol defined in the current study offers a method for generating integration-free iNSCs for the clinical research we developed a novel method for generating integration-free iNSCs. We demonstrated that oriP/EBNA1-based episomal vectors could generate iNSCs by single transfection

2015-03-27 | E-GEOD-67319 | biostudies-arrayexpress

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

| PRJNA141519 | ENA

Therapeutic globin expression in thalassemia patient induced pluripotent stem cells from genomic safe harbors

Project description:The advent of human induced pluripotent stem (iPS) cells enables for the first time the derivation of unlimited numbers of patient-specific stem cells and holds great promise for regenerative medicine. However, realizing the full potential of iPS cells requires robust, precise and safe strategies for their genetic modification. Safe human iPS cell engineering is especially needed for therapeutic applications, as stem cell-based therapies that rely on randomly integrated transgenes pose oncogenic risks. Here we describe a strategy to genetically modify iPS cells from patients with beta-thalassemia in a potentially clinically relevant manner. Our approach is based on the identification and selection of “safe harbor” sites for transgene expression in the human genome. We show that thalassemia patient iPS cell clones harboring a transgene can be isolated and screened according to chromosomal position. We next demonstrate that iPS cell clones that meet our “safe harbor” criteria resist silencing and allow for therapeutic levels of beta-globin expression upon erythroid differentiation without perturbation of neighboring gene expression. Combined bioinformatics and functional analyses thus provide a robust and dependable approach for achieving desirable levels of transgene expression from selected chromosomal loci. This approach may be broadly applicable to introducing therapeutic or suicide genes into patient specific iPS cells for use in cell therapy.

2010-10-26 | GSE24901 | GEO

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