| Usefulness of a non-invasive reporter system for monitoring reprogramming state in pig cells: results of a cell fusion experiment. | |
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MedLine Citation:
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PMID: 20431247 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
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Dedifferentiation of differentiated cells such as fibroblasts into pluripotent stem cells, so-called iPS cells, was first reported by Yamanaka et al., who successfully employed retroviral gene delivery of four stem-cell-specific transcription factors (Oct-3/4, Klf4, Sox2 and c-myc). Despite the mouse system in which an Oct-3/4 or Nanog promoter-based reporter system has already been established, there is no useful system in pigs for reporting the reprogramming state of gene-engineered cells. In this study, we constructed a pOEIN plasmid carrying a ca. 5.4-kb mouse Oct-3/4 promoter linked to the EGFP cDNA and neomycin expression unit and produced a porcine embryonic cell line stably incorporating it in the genome. Cell fusion with mouse embryonal carcinoma cell line F9 resulted in generation of colonies with distinct EGFP-derived fluorescence around 14 days after fusion. RT-PCR using these colonies also confirmed expression of endogenous porcine pluripotency-specific Oct-3/4, Sox2 and Stat3 mRNA. These findings suggest that mouse-derived components are sufficient to induce dedifferentiation of differentiated pig cells and also that reprogramming proceeds gradually. The present non-invasive reporter system will be useful to better define the reprogramming mechanism and/or to identify novel reprogramming molecules in the pig. |
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Authors:
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Akio Ozawa; Eri Akasaka; Satoshi Watanabe; Mitsutoshi Yoshida; Kazuchika Miyoshi; Masahiro Sato |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-04-21 |
Journal Detail:
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Title: The Journal of reproduction and development Volume: 56 ISSN: 0916-8818 ISO Abbreviation: J. Reprod. Dev. Publication Date: 2010 Aug |
Date Detail:
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Created Date: 2010-09-06 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9438792 Medline TA: J Reprod Dev Country: Japan |
Other Details:
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Languages: eng Pagination: 363-9 Citation Subset: IM |
Affiliation:
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Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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