論文

査読有り 国際誌
2018年11月15日

Generation of Footprint-Free Canine Induced Pluripotent Stem Cells Using Auto-Erasable Sendai Virus Vector.

Stem cells and development
  • Masaya Tsukamoto
  • ,
  • Toshiya Nishimura
  • ,
  • Kyohei Yodoe
  • ,
  • Ryoji Kanegi
  • ,
  • Yasunori Tsujimoto
  • ,
  • Md Emtiaj Alam
  • ,
  • Mizuki Kuramochi
  • ,
  • Mitsuru Kuwamura
  • ,
  • Manami Ohtaka
  • ,
  • Ken Nishimura
  • ,
  • Mahito Nakanishi
  • ,
  • Toshio Inaba
  • ,
  • Kikuya Sugiura
  • ,
  • Shingo Hatoya

27
22
開始ページ
1577
終了ページ
1586
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1089/scd.2018.0084

Canine induced pluripotent stem cells (ciPSCs) can be used in regenerative medicine. However, there are no reports on the generation of genome integration-free and completely exogenous gene-silenced (footprint free) ciPSCs that are tolerant to enzymatic single-cell passage. In this study, we reprogrammed canine embryonic fibroblasts using the auto-erasable replication-defective and persistent Sendai virus vector, SeVdp(KOSM)302L, and generated two ciPSC lines. The ciPSCs were positive for pluripotent markers, including alkaline phosphatase activity as well as OCT3/4, SOX2, and NANOG transcripts, and NANOG, stage-specific embryonic antigen-1, and partial TRA-1-60 protein expression, even after SeVdp(KOSM)302L removal. The ciPSCs were induced to differentiate into all the three germ layers as embryoid bodies in vitro and as teratomas in vivo. Furthermore, SeVdp(KOSM)302L-free ciPSCs maintained a normal karyotype even after repeated enzymatic single-cell passaging. Therefore, to our knowledge, for the first time, we demonstrated the generation of footprint-free and high-quality ciPSCs that can be passaged at the single-cell stage using enzymatic methods. Our method for generation of ciPSCs is a good step toward the development of clinical application of ciPSCs.

リンク情報
DOI
https://doi.org/10.1089/scd.2018.0084
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/30215317

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