The isolation of stem cells from the human exfoliated deciduous teeth by genetic engineering

• Project/Area Number: 26463109
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthodontics/Pediatric dentistry
• Research Institution: Niigata University
• Principal Investigator: SAWAMI Tadashi 新潟大学, 医歯学総合研究科, 特任助教 (90710442)
• Co-Investigator(Kenkyū-buntansha): 佐藤 正宏 鹿児島大学, 医用ミニブタ・先端医療開発研究センター, 教授 (30287099) ; 齊藤 一誠 新潟大学, 医歯学系, 准教授 (90404540) ; 野口 洋文 琉球大学, 医学(系)研究科(研究院), 教授 (50378733) ; 稲田 絵美 鹿児島大学, 医歯学域附属病院, 助教 (30448568)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 再生歯学 / 神経幹細胞

Outline of Final Research Achievements

It was considered that to regenerate nerve cells is difficult, but the regeneration of nerve cells from neural stem cells gradually have been showed by the development of a recent study about regenerative medicine.Our aims in this study were to establish the way to isolate stem cells from the human exfoliated deciduous teeth by genetic engineering, and to prove the isolated stem cell effective as of material to tooth regeneration.
We showed that piggy Bac genetic electroporation using Neon was effective to the pulp cell of deciduous teeth and iPS cell. We also isolated dental stem cells from the human exfoliated deciduous teeth by genetic engineering, and showed a neural stem cell marker from the isolated stem cells. The nerve cell was admitted by in vitro differentiation derived from them.

Research Products

• [Journal Article] Isolation and characterization of lymphoid enhancer factor-1-positive deciduous dental pulp stem-like cells after transfection with a piggyBac vector containing LEF1 promoter-driven selection markers (2017)
  • Author(s): Tomoya Murakami, Issei Saitoh, Masahiro Sato, Emi Inada, Miki Soda, Masataka Oda, Hisanori Domone, Yoko Iwase, Tadashi Sawami, Kazunari Matsueda, Yutaka Terao, Hayato Ohshima, Hirofumi Noguchi, Haruaki Hayasaki
  • Journal Title: Archives of Oral Biology Volume: 印刷中
  • Acknowledgement Compliant
• [Journal Article] PiggyBac transposon-mediated gene delivery efficiently generates stable transfectants derived from cultured primary human deciduous tooth dental pulp cells (HDDPCs) and HDDPC-derived iPS cells (2015)
  • Author(s): Inada E, Saitoh I, Miura H, Ohtsuka M, Murakami T, Sawami T, Yamasaki Y, Watanabe S, Aoki R, Sato M
  • Journal Title: Int J Oral Sci Volume: 7(3) Pages: 144-154
• [Presentation] The genetic engineering-based isolation of lymphoid enhancer-binding factor-1 (LEF1) positive stem-like cells from human deciduous tooth cell-derived iPSCs (2016)
  • Author(s): Tomoya Murakami, Issei Saitoh, Masahiro Sato, Emi Inada, Miki Soda, Yoko Iwase, Tadashi Sawami, Ayako Suzuki, Hayato Ohshima, Haruaki hayasaki.
  • Organizer: 10th Biennial Conference of the Pediatric Dentistry Association of Asia in conjunction with 54th Annual Conference of the Japanese Society of Pediatric Dentistry
  • Place of Presentation: 東京ドームホテル(東京都文京区)
  • Year and Date: 2016-05-26
  • Int'l Joint Research
• [Presentation] The genetic engineering-based isolation of lymphoid enhancer-binding factor-1 (LEF1) positive stem-like cells from human deciduous tooth cell-derived iPSCs. (2016)
  • Author(s): Murakami T, Saitoh I, Sato M, Inada E, Soda M, Iwase Y, Sawami T, Suzuki A, Ohshima H,HAYASAKI H
  • Organizer: 45rd Annual Meeting & Exhibition of the AADR
  • Place of Presentation: Los Angeles, USA
  • Year and Date: 2016-03-16
  • Int'l Joint Research
• [Presentation] ALP as a reliable marker for predicting early reprogramming (2016)
  • Author(s): Soda M, Saitoh I, Inada E, Murakami T, Suzuki A, Sawami T, Kagoshima A, Iwase Y, Terao Y, Ohshima H, Hayasaki H, Sato M
  • Organizer: 45rd Annual Meeting & Exhibition of the AADR
  • Place of Presentation: Los Angeles, USA
  • Year and Date: 2016-03-16
  • Int'l Joint Research
• [Presentation] Lymphoid enhancer factor-1 promoterを用いた乳歯歯髄幹細胞様細胞の単離 (2015)
  • Author(s): 村上智哉、齊藤一誠、左右田美樹、澤味 規、鹿児島暁子、寺尾 豊、大島勇人、早崎治明
  • Organizer: 平成27年度新潟歯学会第2回例会
  • Place of Presentation: 新潟大学歯学部（新潟県・新潟市）
  • Year and Date: 2015-11-07
• [Presentation] ヒト乳歯歯髄細胞のアルカリホスファターゼ活性とOCT-3/4発現はiPS細胞樹立の可否を予測する有効なマーカーである (2015)
  • Author(s): 稲田絵美，佐藤正宏，齊藤一誠，窪田直子，澤味 規，村上智哉，左右田美樹，早崎治明，山﨑要一
  • Organizer: 第53回日本小児歯科学会大会
  • Place of Presentation: 広島国際会議場（広島県・広島市）
  • Year and Date: 2015-05-21
• [Presentation] PiggyBac-transposon-mediated Gene-delivery Efficiently Generates Stable Transfectants From HDDPCs and HDDPC-derived-iPSCs (2015)
  • Author(s): M. SODA, I. SAITOH, E. INADA, T. MURAKAMI, Y. IWASE, N. KUBOTA, T. SAWAMI, Y. MATSUMOTO, Y. YAMASAKI, H. HAYASAKI, H. OHSHIMA, M. SATO
  • Organizer: International Association for Dental Reseach
  • Place of Presentation: John B. Hynes Veterans Memorial Convention Center,Boston, Mass., USA
  • Year and Date: 2015-03-11 – 2015-03-15