Generation of induced pluripotent stem (iPS) cells using non-viral magnet-based nanofection
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE21357 | GEO | 2024/02/29
REPOSITORIES: GEO
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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.
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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.
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Project description:Whole-genome expression analysis of non-integrating Sendai virus (SeV) vector and retroviral vector-generated human iPS cells. Parental fibroblast and induced pluripotent stem (iPS) cells.
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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).
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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. 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 using the RMA package of Bioconductor.
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Project description:iPS-OSH was the novel iPS cells generated from our lab, the microarray analysis was used for determining the gene expression profiles of iPS-OSH. iPS-J was provided from Dr. Yamanaka (Riken, APS0001). Microarray analysis was used to compare the difference of gene expression profiles in iPS-OSH, iPS-J, ESC and MEF.
2012-08-01 | GSE31822 | GEO
Project description:iPS-OSH was the novel iPS cells generated from our lab, the microarray analysis was used for determining the gene expression profiles of iPS-OSH. iPS-J was provided from Dr. Yamanaka (Riken, APS0001). Microarray analysis was used to compare the difference of gene expression profiles in iPS-OSH, iPS-J, ESC and MEF. The total RNA extracted from iPS-OSH, iPS-J, ESC and MEF was used for the microarray analysis to compare the gene expression profiles.
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Project description:We present a robust serum-free system for the rapid and efficient reprogramming of mouse somatic cells by Oct4, Sox2 and Klf4. The elimination of fetal bovine serum and oncogene c-Myc allowed reprogramming cells to be detected as early as Day 2 and reached greater than 10% of the population at Day 7 post retroviral transduction. The resulting iPS colonies were isolated with high efficiency to establish pluripotent cell lines. Based on this method, we further developed iPS-SF1 as a dedicated reprogramming medium for chemical screening and mechanistic investigations. Comparasion of iPS cell lines derived from serum-free culture condition, MEF cells cultured in serum and serum-free condition and ES cell lines was shown. Experiment Overall Design: Selected induced pluripotent stem cell lines and its original mouse embroynic fibroblast were comparaed to defined mouse embroynic stem cell lines: R1 and CGR8.
2009-03-28 | E-GEOD-15267 | biostudies-arrayexpress
Project description:Transfer of somatic cell nuclei to enucleated eggs or ectopic expression of specific transcription factors are two different reprogramming strategies used to generate pluripotent cells from differentiated cells. However, they are poorly efficient and other unknown factors might be required to increase their success rate. Here, we show that Xenopus egg extracts at the metaphase stage (M phase) have a strong reprogramming activity on mouse embryonic fibroblasts (MEFs). First, they reset replication properties of MEF nuclei toward a replication profile characteristic of early development and they erase several epigenetic marks, such as trimethylation of H3K9, H3K4 and H4K20. Second, when MEFs are reversibly permeabilized in the presence of M phase Xenopus egg extracts, they show a transient increase in cell proliferation, form colonies and start to express specific pluripotency markers. Finally, transient exposure of MEF nuclei to M phase Xenopus egg extracts increases the success of nuclear transfer to enucleated mouse oocytes and strongly synergize with the production of pluripotent stem cells by ectopic expression of transcription factors. The mitotic stage of the egg extract is crucial as neither of these effects is detected when using interphasic Xenopus egg extracts. Our data demonstrate that mitosis is essential to make mammalian somatic nuclei prone to reprogramming and that, surprisingly, the heterologous Xenopus system has features that are conserved enough to remodel mammalian nuclei. MEF cells were infected by retroviruses encoding for Oct4, Sox2, Klf4 and c-Myc and, then reversibly permeabilized and treated with Xenopus M-phase extract. All samples of each cell type (e.g. untreated MEF, ES cells and M-iPS cells) were made in triplicates. Total RNAs were extracted with RNeasy kit and hybridized on Nimblegen MM9 microarrays.
2011-11-21 | E-GEOD-31691 | biostudies-arrayexpress
Project description:We present a robust serum-free system for the rapid and efficient reprogramming of mouse somatic cells by Oct4, Sox2 and Klf4. The elimination of fetal bovine serum and oncogene c-Myc allowed reprogramming cells to be detected as early as Day 2 and reached greater than 10% of the population at Day 7 post retroviral transduction. The resulting iPS colonies were isolated with high efficiency to establish pluripotent cell lines. Based on this method, we further developed iPS-SF1 as a dedicated reprogramming medium for chemical screening and mechanistic investigations. Comparasion of iPS cell lines derived from serum-free culture condition, MEF cells cultured in serum and serum-free condition and ES cell lines was shown.
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