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DNA damage responses in human induced pluripotent stem cells and embryonic stem cells

Momcilovic, O and Knobloch, L and Fornsaglio, J and Varum, S and Easley, C and Schatten, G (2010) DNA damage responses in human induced pluripotent stem cells and embryonic stem cells. PLoS ONE, 5 (10).

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Abstract

Background: Induced pluripotent stem (iPS) cells have the capability to undergo self-renewal and differentiation into all somatic cell types. Since they can be produced through somatic cell reprogramming, which uses a defined set of transcription factors, iPS cells represent important sources of patient-specific cells for clinical applications. However, before these cells can be used in therapeutic designs, it is essential to understand their genetic stability. Methodology/Principal Findings: Here, we describe DNA damage responses in human iPS cells. We observe hypersensitivity to DNA damaging agents resulting in rapid induction of apoptosis after γ-irradiation. Expression of pluripotency factors does not appear to be diminished after irradiation in iPS cells. Following irradiation, iPS cells activate checkpoint signaling, evidenced by phosphorylation of ATM, NBS1, CHEK2, and TP53, localization of ATM to the double strand breaks (DSB), and localization of TP53 to the nucleus of NANOG-positive cells. We demonstrate that iPS cells temporary arrest cell cycle progression in the G phase of the cell cycle, displaying a lack of the G /S cell cycle arrest similar to human embryonic stem (ES) cells. Furthermore, both cell types remove DSB within six hours of γ-irradiation, form RAD51 foci and exhibit sister chromatid exchanges suggesting homologous recombination repair. Finally, we report elevated expression of genes involved in DNA damage signaling, checkpoint function, and repair of various types of DNA lesions in ES and iPS cells relative to their differentiated counterparts. Conclusions/Significance: High degrees of similarity in DNA damage responses between ES and iPS cells were found. Even though reprogramming did not alter checkpoint signaling following DNA damage, dramatic changes in cell cycle structure, including a high percentage of cells in the S phase, increased radiosensitivity and loss of DNA damage-induced G /S cell cycle arrest, were observed in stem cells generated by induced pluripotency. © 2010 Momcilovic et al. 2 1 1


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Momcilovic, O
Knobloch, L
Fornsaglio, J
Varum, S
Easley, C
Schatten, Ggps15@pitt.eduGPS15
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorCapogrossi, Maurizio C.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Date: 17 November 2010
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 5
Number: 10
DOI or Unique Handle: 10.1371/journal.pone.0013410
Schools and Programs: Graduate School of Public Health > Human Genetics
Refereed: Yes
MeSH Headings: Blotting, Western; Cell Cycle; Cell Cycle Proteins--metabolism; Cell Differentiation; Cell Line; DNA Damage; DNA Repair; Embryonic Stem Cells--cytology; Embryonic Stem Cells--metabolism; Flow Cytometry; Gene Expression Profiling; Humans; Immunohistochemistry; Microscopy, Confocal; Phosphorylation; Pluripotent Stem Cells--cytology; Pluripotent Stem Cells--metabolism; Polymerase Chain Reaction
Other ID: NLM PMC2955528
PubMed Central ID: PMC2955528
PubMed ID: 20976220
Date Deposited: 22 Aug 2012 21:03
Last Modified: 30 Mar 2021 15:55
URI: http://d-scholarship.pitt.edu/id/eprint/13561

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