Preliminary study of siRNA transfection into embryonic lingual tissue to establish the technique for lingual papillae regeneration

• Project/Area Number: 25463153
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Surgical dentistry
• Research Institution: The Nippon Dental University
• Principal Investigator: Yoshimura Ken 日本歯科大学, 新潟生命歯学部, 准教授 (90297953)
• Co-Investigator(Renkei-kenkyūsha): NASHIDA TOMOKO 日本歯科大学, 新潟生命歯学部, 准教授 (10133464) ; MIKAMI MASATO 日本歯科大学, 新潟生命歯学部, 講師 (90173997)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 遺伝子導入 / siRNA / 舌乳頭形態形成 / 舌 / 形態形成 / 舌粘膜 / Electropolation / 胎仔

Outline of Final Research Achievements

Essential proteins for lingual papillae morphogenesis are likely present during embryonic development. We searched for the responsible genes and selected candidates involved in lingual papillae morphogenesis. Then to establish regeneration techniques, preliminary transfection of siRNA was performed. Transfection of the synthesized siRNA together with vectors for fluorescent proteins expression into rat embryonic lingual tissues was performed using the electroporation method. After the transfection, the lingual tissues were organ-cultured and observed. Twenty-four hours after the transfection, fluorescence markers started illuminating inside of the lingual tissues and were continuing to glow up to the 72 hour intervals. However, the quality of fluorescence was being occasionally unstable. Optimization of electric pulse parameters was required in order to maintain cell viability and more efficacious transfection methods also would be required.

Research Products

• [Presentation] Morphological evaluation of transfection in rat embryonic organ-cultured lingual tissue by the in vitro electroporation method (2015)
  • Author(s): Ken Yoshimura,Tomoko Nashida,Masato Mikami,Ikuo Kageyama
  • Organizer: 日本歯科大学歯学会
  • Place of Presentation: 日本歯科大学 新潟生命歯学部 アイヴイホール
  • Year and Date: 2015-06-06
• [Presentation] An evaluation of transfection in rat embryonic organ-cultured lingual tissue by the in vitro electroporation method． (2014)
  • Author(s): Yoshimura K, Nashida T, Mikami M and Kageyama I
  • Organizer: 歯科基礎医学会
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2014-09-25 – 2014-09-27