Role of p53 in mouse iPS cell generation
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE13312 | GEO | 2009/03/01
SECONDARY ACCESSION(S): PRJNA113973
REPOSITORIES: GEO
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Project description:p53 knockdown by shRNA markedly increased efficiency of human iPS cell generation Gene expression patterns were compared between human ES cells and dermal fibroblasts. Gene expression patterns were also compared between p53 shRNA-treated fibroblasts and control fibroblasts.
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Project description:We observed a marked increase in efficiency of iPS cell generation when p53 is deleted. Experiment Overall Design: Gene expression patterns were compared between p53 wt MEF and p53-null MEF cells. Experiment Overall Design: Gene expression patterns were also compared between ES cells and MEF.
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Project description:This SuperSeries is composed of the following subset Series: GSE13312: Role of p53 in mouse iPS cell generation GSE13334: Effect of p53 in human iPS cell generation Refer to individual Series
2009-02-28 | E-GEOD-13365 | biostudies-arrayexpress
Project description:Human fibroblasts can be induced into pluripotent stem cells (iPS cells), but the reprogramming efficiency is quite low. Here, we screened a panel of candidate factors in the presence of OCT4, SOX2, KLF4 and c-MYC in an effort to improve the reprogramming efficiency from human adult fibroblasts. We found that p53 siRNA and UTF1 enhanced the efficiency of iPS cell generation up to 100-fold, even when the oncogene c-MYC was removed from the combinations. We further demonstrated that by using a novel combination of the four factors OCT4, SOX2, KLF4 and UTF1, iPS cells could be generated at a frequency at least 10 times higher than using the original four reprogramming factors without c-MYC. The iPS cells generated in this work have a similar gene expression profile and differentiation potential as human embryonic stem (hES) cells. In conclusion, two novel supporting factors that increase the efficiency of direct reprogramming have been identified, and a more-efficient method for the generation of human iPS cells has been developed in the absence of the oncogene c-MYC. Keywords: cell type comparison
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Project description:Human fibroblasts can be induced into pluripotent stem cells (iPS cells), but the reprogramming efficiency is quite low. Here, we screened a panel of candidate factors in the presence of OCT4, SOX2, KLF4 and c-MYC in an effort to improve the reprogramming efficiency from human adult fibroblasts. We found that p53 siRNA and UTF1 enhanced the efficiency of iPS cell generation up to 100-fold, even when the oncogene c-MYC was removed from the combinations. We further demonstrated that by using a novel combination of the four factors OCT4, SOX2, KLF4 and UTF1, iPS cells could be generated at a frequency at least 10 times higher than using the original four reprogramming factors without c-MYC. The iPS cells generated in this work have a similar gene expression profile and differentiation potential as human embryonic stem (hES) cells. In conclusion, two novel supporting factors that increase the efficiency of direct reprogramming have been identified, and a more-efficient method for the generation of human iPS cells has been developed in the absence of the oncogene c-MYC. Keywords: cell type comparison Total RNA from hFSF, hAFF, hES cells (H1, H7) and 7 established iPS cell lines were labeled with Cy5, hybridized to a human Oligo Microarray (Phalanx Human Whole Genome OneArray™, Phalanx Biotech) according to the manufacturer's protocol. Three technical repetitions were performed. After hybridization, arrays were scanned using GenePix 4000B scanner (Molecular Devices) and processed using the GenePix Pro 6.0 software (Molecular Devices). After removing control probes, a 14/33 presence call (SNR>=5 and foreground-background>0) was used to filter probes for the 33 microarrays, resulting in 12311 probes for further quantile normalization.
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Project description:The primary aim of this study is to evaluate the effect of transient knock down of P53 as a tool to increase the efficiency of a non-integrative methodology for reprogramming adult human normal dermal fibroblasts. This study demonstrate that transient knockdown of P53 is an efficient way to produce iPSC containing minimal genomic alterations, which meets the increased demand for iPSC in personalized drug screening campaigns. Total RNA was isolated from 3 iPS cell lines generated without P53 knockdown and 3 generated with P53 knockdown. In addition total RNA was isolated from the parental normal human dermal fibroblasts and from a reference human iPS cell line from Systembio (SBI).
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Project description:In this study, we sought to examine whether an extracellular matrix (ECM)-based xeno-free culture system that we recently established could be used together with a microRNA-enhanced mRNA reprogramming method for the generation of clinically safe iPS cells. The notable features of this method are (1) the use of a xeno-free/feeder-free culture system for the generation and expansion of iPS cells rather than the conventional labor-intensive culture systems using human feeder cells or human feeder-conditioned medium and (2) the enhancement of mRNA-mediated reprogramming via the delivery of microRNAs. Strikingly, we observed the early appearance of iPS cell colonies (~11 days), substantial reprogramming efficiency (~0.2-0.3%), and a high percentage of ESC-like colonies among the total colonies (~87.5%), indicating enhanced kinetics and reprogramming efficiency. Therefore, the combined method established in this study provides a valuable platform for the generation and expansion of clinically safe (i.e., integration- and xeno-free) iPS cells, facilitating immune-matched cell therapy in the near future.
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Project description:Using magneto-nanofection, iPS cells were efficiently generated by the transient expression of iPS genes in MEF (mouse embryonic fibroblast), suggesting that the non-viral magneto-nanofection method can be used for the efficient generation of iPS cells.
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