Establishment of safe and efficient differentiation system using automatic removal vector

2013 Fiscal Year Final Research Report

• Project/Area Number: 24700474
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biomedical engineering/Biological material science
• Research Institution: University of Tsukuba
• Principal Investigator: NISHIMURA Ken 筑波大学, 医学医療系, 助教 (80500610)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: 分化 / 多能性幹細胞 / 残存未分化細胞 / SeVdpベクター / miRNA

Research Abstract

When we induce differentiated cells from pluripotent stem cells, the risks of tumor formation by remained undifferentiated cells must be concerned. To reduce such a risk, we tried to establish a system for elimination of undifferentiated cells in the differentiated tissue using our SeVdp vector system. The SeVdp vector which has the target sequence of neuron-specific miRNA was infected to mouse iPS cells, and then they were differentiated to neuron. As a result, the vector were automatically removed from neuron while it was persistently infected in undifferentiated cells. But when we tried to select differentiated cells using negative selection gene from the vector, not only undifferentiated cells but also differentiated cells were killed by the treatment. We plan to investigate other method of differentiation and negative selection to selectively eliminate the undifferentiated cells.

Research Products

• [Journal Article] Heterogeneous Nuclear Ribonucleoprotein R Cooperates with Mediator to Facilitate Transcription Reinitiation on the c-Fos Gene (2013)
  • Author(s): A. Fukuda, M. Shimada, T. Nakadai, K. Nishimura, K. Hisatake
  • Journal Title: PLOS ONE Volume: Vol. 8
  • DOI: 10.1371/journal.pone.0072496
  • Peer Reviewed
• [Journal Article] GPAT2, a mitochondrial outer membrane protein, in piRNA biogenesis in germline stem cells (2013)
  • Author(s): Y. Shiromoto, S. Kuramochi-Miyagawa, A. Daiba, S. Chuma, A. Katanaya, A. Katsumata, K. Nishimura, M. Ohtaka, M. Nakanishi, T. Nakamura, K. Yoshinaga, N. Asada, S. Nakamura, T. Yasunaga, K. Kojima-Kita, D. Itou, T. Kimura, T. Nakano
  • Journal Title: RNA Volume: Vol. 19 Pages: 803-810
  • Peer Reviewed
• [Journal Article] Expansion of Functional Human Mucosal-Associated Invariant T Cells via Reprogramming to Pluripotency and Redifferentiation (2013)
  • Author(s): H. Wakao, K. Yoshikiyo, U. Koshimizu, T. Furukawa, K. Enomoto, T. Matsunaga, T. Tanaka, Y. Yasutomi, T. Yamada, H. Minakami, J. Tanaka, A. Oda, T. Sasaki, R. Wakao, O. Lantz, T. Udagawa, Y. Sekiya, K. Higuchi, N. Harada, K. Nishimura, M. Ohtaka, M. Nakanishi, H. Fujita
  • Journal Title: Cell Stem Cells Volume: Vol. 12 Pages: 546-558
  • Peer Reviewed
• [Journal Article] Generation of rejuvenated antigen-specific T cells by reprogramming to pluripotency and redifferentiation (2013)
  • Author(s): T. Nishimura, S. Kaneko, A. Kawana-Tachikawa, Y. Tajima, H. Goto, D. Zhu, K. Nakayama-Hosoya, S. Iriguchi, Y. Uemura, T. Shimizu, N. Takayama, D. Yamada, K. Nishimura, M. Ohtaka, N. Watanabe, S. Takahashi, A. Iwamoto, H. Koseki, M. Nakanishi, K. Eto, H. Nakauchi
  • Journal Title: Cell Stem Cells Volume: Vol. 12 Pages: 114-126
  • Peer Reviewed
• [Journal Article] Podocalyxin is a Glycoprotein Ligand of the Human Pluripotent Stem Cell-Specific Probe rBC2LCN (2013)
  • Author(s): H. Tateno, A. Matsushima, K. Hiemori, Y. Onuma, Y. Ito, K. Hasehira, K. Nishimura, M. Ohtaka, S. Takayasu, M. Nakanishi, Y. Ikehara, M. Nakanishi, K. Ohnuma, T. Chan, M. Toyoda, H. Akutsu, A. Umezawa, M. Asashima, J. Hirabayashi
  • Journal Title: Stem Cells Trans. Med. Volume: Vol. 2 Pages: 265-273
  • Peer Reviewed
• [Presentation] 初期化が途中で停止した細胞を用いたiPS細胞誘導過程の解析 (2014)
  • Author(s): 西村健
  • Organizer: 第13回日本再生医療学会総会
  • Place of Presentation: 京都国際会議場、京都
  • Year and Date: 2014-03-05
• [Presentation] Klf4遺伝子発現量調節による初期化が途中で停止した細胞の誘導 (2013)
  • Author(s): 西村健
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸国際展示場、神戸
  • Year and Date: 2013-12-03
• [Presentation] Manipulation of Klf4 expression produces snap shots of iPSC induction (2013)
  • Author(s): K. Nishimura
  • Organizer: CSHA/ISSCR Joint Meeting on Stem Cells in Science and Medicine
  • Place of Presentation: Suzhou Dushu Lake Conference Center、Suzhou、China
  • Year and Date: 2013-10-16
• [Presentation] Taking snap shots during iPSC production by SeVdp vectors (2013)
  • Author(s): K. Nishimura
  • Organizer: International Society of Stem Cell Research 11^<th> Annual Meeting
  • Place of Presentation: The Boston Convention Center、Boston、USA
  • Year and Date: 2013-06-14
• [Presentation] SeVdpベクターを用いたiPS細胞の品質調節メカニズムの解析 (2013)
  • Author(s): 西村健
  • Organizer: 第12回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2013-03-22
• [Presentation] Application of Cytoplasmic RNA vector Developed from Noncytopathic RNA Viruses to Stem Cell Engineering (2013)
  • Author(s): K. Nishimura
  • Organizer: The 2nd Meeting on RNA and Biofunctions-Asia Study
  • Place of Presentation: ホテルレオパレス福岡、福岡
  • Year and Date: 2013-01-09
• [Presentation] QUALITY CONTROL OF IPS CELLS BY UNIQUE GENE TRANSFER SYSTEM (2012)
  • Author(s): K. Nishimura
  • Organizer: International Society of Stem Cell Research 10^<th> Annual Meeting
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2012-06-14