Pluripotent cell derivation from male germline cells by suppression of <i>Dmrt1</i> and <i>Trp53</i>

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  • TANAKA Takashi
    Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan
  • KANATSU-SHINOHARA Mito
    Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan Japan Science and Technology Agency, PRESTO, Kyoto 606-8501, Japan
  • HIROSE Michiko
    The Institute for Physical and Chemical Research (RIKEN), Bioresource Center, Tsukuba 305-0074, Japan
  • OGURA Atsuo
    The Institute for Physical and Chemical Research (RIKEN), Bioresource Center, Tsukuba 305-0074, Japan
  • SHINOHARA Takashi
    Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan

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  • Pluripotent cell derivation from male germline cells by suppression of Dmrt1 and Trp53

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Abstract

Diploid germ cells are thought to have pluripotency potential. We recently described a method to derive pluripotent stem cells (PSCs) from cultured spermatogonial stem cells (SSCs) by depleting Trp53 and Dmrt1, both of which are known suppressors of teratomas. In this study, we used this technique to analyze the effect of this protocol in deriving PSCs from the male germline at different developmental stages. We collected primordial germ cells (PGCs), gonocytes and spermatogonia, and the cells were transduced with lentiviruses expressing short hairpin RNA against Dmrt1 and/or Trp53. We found that PGCs are highly susceptible to reprogramming induction and that only Trp53 depletion was sufficient to induce pluripotency. In contrast, gonocytes and spermatogonia were resistant to reprogramming by double knockdown of Dmrt1 and Trp53. PSCs derived from PGCs contributed to chimeras produced by blastocyst injection, but some of the embryos showed placenta-only phenotypes suggestive of epigenetic abnormalities of PGC-derived PSCs. These results show that PGCs and gonocytes/spermatogonia have distinct reprogramming potential and also suggest that fresh and cultured SSCs do not necessarily have the same properties.

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