Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells

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Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells

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Title: Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells
Author: Kang, Hoon-Chul; Yang, Eungi; Park, Kyung-Soon; Lee, Kyung-Ah; Hwang, Dong-Youn; Lanza, Robert; Reh, Thomas A.; Chang, Mi-Yoon; Kim, Chun-Hyung; Kim, Dohoon; Moon, Jung-Il; Ko, Sanghyeok; Park, Junpil; Chung, Young; Kim, Kwang-Soo

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Citation: Chang, Mi-Yoon, Dohoon Kim, Chun-Hyung Kim, Hoon-Chul Kang, Eungi Yang, Jung-Il Moon, Sanghyeok Ko, et al. 2010. Direct reprogramming of rat neural precursor cells and fibroblasts into pluripotent stem cells. PLoS ONE 5(3): e9838.
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Abstract: Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem (iPS) cells would provide researchers with a powerful tool for studying human physiology and disease. Here, we report direct reprogramming of rat neural precursor (NP) cells and rat embryonic fibroblasts (REF) into iPS cells by retroviral transduction using either three (Oct3/4, Sox2, and Klf4), four (Oct3/4, Sox2, Klf4, and c-Myc), or five (Oct3/4, Sox2, Klf4, c-Myc, and Nanog) genes. Methodology and Principal Findings: iPS cells were generated from both NP and REF using only three (Oct3/4, Sox2, and Klf4) genes without c-Myc. Two factors were found to be critical for efficient derivation and maintenance of rat iPS cells: the use of rat instead of mouse feeders, and the use of small molecules specifically inhibiting mitogen-activated protein kinase and glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming, but failed to generate iPS cells. However, when combined with retroviral transduction, this method generated iPS cells with significantly higher efficiency. Morphology, gene expression, and epigenetic status confirmed that these rat iPS cells exhibited ESC-like properties, including the ability to differentiate into all three germ layers both in vitro and in teratomas. In particular, we found that these rat iPS cells could differentiate to midbrain-like dopamine neurons with a high efficiency. Conclusions/Significance: Given the usefulness of rats as an experimental system, our optimized method would be useful for generating rat iPS cells from diverse tissues and provide researchers with a powerful tool for studying human physiology and disease.
Published Version: doi:10.1371/journal.pone.0009838
Other Sources: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2844422/pdf/
Terms of Use: This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA
Citable link to this page: http://nrs.harvard.edu/urn-3:HUL.InstRepos:4621704

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