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Generation of CD44 gene-deficient mouse derived induced pluripotent stem cells

CD44 gene-deficient iPSCs

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Abstract

Induced pluripotent stem cells (iPSCs) show good promise for the treatment of defects caused by numerous genetic diseases. Herein, we successfully generated CD44 gene-deficient iPSCs using Oct4, Sox2, Klf4, and vitamin C. The generated iPSCs displayed a characteristic morphology similar to the well-characterized embryonic stem cells. Alkaline phosphatase, cell surface (SSEA1, NANOG, and OCT4), and pluripotency markers were expressed at high levels in these cells. The iPSCs formed teratomas in vivo and supported full-term development of constructed porcine embryos by inter-species nuclear transplantation. Importantly, incubation with trichostatin A increased the efficiency of iPSCs generation by increasing the histone acetylation levels. Moreover, more iPSCs colonies appeared following cell passaging during colony picking, thus increasing the effectiveness of iPSCs selection. Thus, our work provides essential stem cell materials for the treatment of genetic diseases and proposes a novel strategy to enhance the efficiency of induced reprogramming.

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Acknowledgments

This study was funded by the 973 Program (no. 2010CB945404), the National Transgenic Major Program of China (no. 2011ZX08006-005), and “the Fundamental Research Funds for the Central Universities” (no. 11lgpy46).

Conflict of interest

The authors declare that there is no conflict of interest.

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Authors and Affiliations

  1. State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Higher Education Mega Center, Guangzhou, 510006, People’s Republic of China

    Zhenwei Song, Qianqian Ji, Haijing Zhao, Yu Nie, Zuyong He, Yaosheng Chen & Peiqing Cong

Authors
  1. Zhenwei Song
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  2. Qianqian Ji
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  3. Haijing Zhao
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  4. Yu Nie
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  5. Zuyong He
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  6. Yaosheng Chen
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  7. Peiqing Cong
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Corresponding author

Correspondence to Peiqing Cong.

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Editor: T. Okamoto

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Supplementary Table 1

Primers used in this study (DOC 46 kb)

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Song, Z., Ji, Q., Zhao, H. et al. Generation of CD44 gene-deficient mouse derived induced pluripotent stem cells. In Vitro Cell.Dev.Biol.-Animal 50, 874–882 (2014). https://doi.org/10.1007/s11626-014-9786-6

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  • DOI: https://doi.org/10.1007/s11626-014-9786-6

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