Development of hepatocyte induction methods derived from dental pulp and cell transplantation therapy

2013 Fiscal Year Final Research Report

• Project/Area Number: 24792042
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Conservative dentistry
• Research Institution: The Nippon Dental University
• Principal Investigator: TOMINAGA Noriko 日本歯科大学, 生命歯学部, 助教 (90546532)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: 歯髄由来肝細胞 / コロニアルクローニング / 歯髄組織 / αfetoprotein / albumin / human mitochondoria / ETF / SKGII

Research Abstract

Embryotrophic factors (ETFs) derived from squamous cell carcinoma of the cervix (SKG2) were used to induce a hepatocyte population from dental pulp and we isolated. Our aim is to develop new regenerative medicines transplanting established hepatocytes into the damaged liver of animal models, such as into animals with cirrhosis of the liver, and then verifying the recovery of liver function.
We used ETFs for hepatocyte induction, but the ETFs are inactivated prior to use, consequently, during the first year of the research project, we made ETFs collecting from SKG2 conditioned medium by culture in a serum-free medium. In addition, we transplanted hepatocytes from human dental pulp into SCID mice and observed the presence of hepatocytes after transplantation. Immunostaining of the transplanted hepatocytes showed that they expressed a-fetoprotein and albumin, and contained human mitochondria. In addition, the hepatocytes did not become tumorous but did form clusters with the host cells.

Research Products

• [Journal Article] Isolation and characterization of 5mm 300μm 300μm 300μm 300μm 100μm epithelial and myogenic cells by "fishing" for the morphologically distinct cell types in rat primary periodontal ligament culture (2013)
  • Author(s): Tominaga N, Nakahara T, Nasu M, Satoh T
  • Journal Title: Differentiation Volume: volume 85 issue3 Pages: 91-100
• [Journal Article] Isolation and characterization of vascular endothelial cells derived from fetal tooth buds of miniature swine (2013)
  • Author(s): Nasu M, Nakahara T, Tominaga N, Tamaki Y, Ide Y, Tachibana T, Ishikawa H
  • Journal Title: In Vitro Cellular & Developmental Biology - Animal Volume: 49(3) Pages: 189-195
• [Journal Article] Postnatal mandibular cheek tooth development in the miniature pig based on two-dimensional and three-dimensional X-ray analyses (2013)
  • Author(s): Ide Y, Nakahara T, Nasu M, Matsunaga S, Iwanaga T, Tominaga N, Tamaki Y
  • Journal Title: Anat Rec (Hoboken) Volume: 296(8) Pages: 1247-1254
• [Journal Article] ミニブタ胎仔乳臼歯の歯冠由来エナメル上皮細胞の分離培養と遺伝子発現の解析 (2013)
  • Author(s): 中原 貴, 富永徳子, 石川 博
  • Journal Title: 日本口腔外科学会雑誌 Volume: 59 Pages: 266
• [Presentation] Identification of putative stem cells in primary cultures derived from rat molar periodontal ligament and subcultures of myogenic-lineage committed cells (2013)
  • Author(s): Nakahara T, Tominaga N, Ishikawa H
  • Organizer: 11th Annual Meeting International Society for Stem Cell Research (ISSCR)
  • Place of Presentation: Boston Convention and Exhibition Center, Boston, Massachusetts, USA
  • Year and Date: 20130612-15
• [Presentation] Periodontal ligament-derived skeletal muscle cells : isolation from primary whole-tooth cultures by using the "cell-fishing method" (2013)
  • Author(s): Tominaga N, Nakahara T
  • Organizer: The 2nd Trilateral Dental Symposium on "Dental Virtual Education Today"
  • Place of Presentation: The Nippon Dental University, School of Life Dentistry at Tokyo, Kudan hall, Tokyo
  • Year and Date: 2013-11-14
• [Presentation] ラット臼歯歯根膜由来の初代培養および歯根膜由来骨格筋細胞群に幹細胞は存在するか? (2013)
  • Author(s): 富永徳子, 中原 貴, 石川 博
  • Organizer: 第31回日本ヒト細胞学会学術集会
  • Place of Presentation: 所沢市民文化センター「ミューズ」第2 展示室, 所沢
  • Year and Date: 2013-08-10
• [Presentation] クラウン系ミニブタ胎仔の乳臼歯から分離培養したエナメル上皮細胞の遺伝子発現解 (2012)
  • Author(s): 中原 貴, 富永徳子, 石川 博
  • Organizer: 第57回日本口腔外科学会総会・学術大会, パシフィコ横浜
  • Place of Presentation: 横浜
  • Year and Date: 20121019-21
• [Presentation] マウス歯根形成機における頭部エックス線照射による障害歯根の形態およびヘルトヴィッヒ上皮鞘と周囲間葉の細胞動態の観察 (2012)
  • Author(s): 井出吉昭, 中原 貴, 那須優則, 富永徳子, 田巻友一, 石川 博
  • Organizer: 第54回歯科基礎医学会学術大会・総会
  • Place of Presentation: 奥羽大学, 福島
  • Year and Date: 20120914-16
• [Presentation] ラット歯根膜から分離した筋細胞集団の解析 (2012)
  • Author(s): 富永徳子, 中原 貴, 石川 博
  • Organizer: 第10回日本再生歯科医学会学術大会・総会
  • Place of Presentation: ニチイ学館神戸ポートアイランドセンター, 神戸
  • Year and Date: 20120901-02
• [Presentation] 器官再生法で形成された歯根・歯周組織ユニットのマイクロCT を用いた三次元形態解析 (2012)
  • Author(s): 中原 貴, 井出吉昭, 田巻友一, 富永徳子, 佐藤聡, 石川 博
  • Organizer: 第12回日本抗加齢医学会総会
  • Place of Presentation: パシフィコ横浜会議センター, 横浜
  • Year and Date: 20120622-24
• [Presentation] Isolation and characterization of neurons and myogenic cells from rat molar periodontal ligament (2012)
  • Author(s): Tominaga N, Nakahara T, Ishikawa H
  • Organizer: 10th Annual Meeting International Society for Stem Cell Research (ISSCR)
  • Place of Presentation: パシフィコ横浜, 横浜
  • Year and Date: 20120613-16
• [Presentation] ミニブタ胎仔乳臼歯由来エナメル上皮細胞の分離と同定 (2012)
  • Author(s): 中原 貴, 富永徳子, 石川 博
  • Organizer: 第11回日本再生医療学会
  • Place of Presentation: パシフィコ横浜, 横浜
  • Year and Date: 20120612-14
• [Presentation] 再生医療のための骨組織形成-ヒト脂肪組織由来幹細胞を細胞源として- (2012)
  • Author(s): 大山晃弘, 井出吉昭, 田巻友一, 富永徳子, 中原貴, 立花利公, 渡邊美隆, 栗原邦弘, 石川 博
  • Organizer: 第11回日本再生医療学会
  • Place of Presentation: パシフィコ横浜, 横浜
  • Year and Date: 20120612-14
• [Presentation] ヒト脂肪組織由来幹細胞の骨細胞への分化と骨形成 (2012)
  • Author(s): 大山晃弘, 井出吉昭, Tansriratanawong Kallapat, 田巻友一, 富永徳子, 中原 貴, 立花利公, 渡邊美隆, 栗原邦弘, 佐藤 聡, 石川 博
  • Organizer: 平成24年度日本歯科大学歯学会大会, 日本歯科大学生命歯学部メモリアルホール
  • Place of Presentation: 東京
  • Year and Date: 2012-06-02
• [Presentation] 幹細胞医学に基づいた抗加齢医療アプローチの確立に向けて~幹細胞の発生学的起源と抗加齢ポテンシャル (2012)
  • Author(s): 田巻友一, 富永徳子, 井出吉昭, 中原貴
  • Organizer: 平成24年度日本歯科大学歯学会大会, 日本歯科大学生命歯学部メモリアルホール
  • Place of Presentation: 東京
  • Year and Date: 2012-06-02