Fabrication and transplantation of xeno-free canine corneal epithelial cell sheet

• Project/Area Number: 26450422
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: The University of Tokyo
• Principal Investigator: Tsuzuki Keiko 東京大学, 大学院農学生命科学研究科(農学部), 特任助教 (30364251)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 角膜上皮再生 / 犬 / 角膜上皮細胞シート / 犬角膜上皮細胞 / 角膜上皮シート / 角膜移植 / 角膜再生 / 犬角膜上皮細胞シート / 角膜損傷 / 犬角膜上皮細胞株 / 再生医療

Outline of Final Research Achievements

It was revealed that canine corneal epithelial cells can maintain their undifferentiated state and proliferation ability even when cultured without any feeder cells. Thus, feeder cells can be eliminated for fabrication of canine corneal epithelial cell sheets. However, canine corneal epithelial cells showed apparent morphological change when cultured using autologous serum. Therefore, the use of fetal bovine serum could not be eliminated to fabricate canine corneal epithelial cell sheets. In the process to explore the possible growth factors to fabricate canine corneal epithelial cells, we found out that hepatocyte growth factor (HGF) promotes the cell invasion ability of canine corneal epithelial cells and HGF can be a promising drug to treat corneal injury. We could also developed transplantation of canine corneal epithelial cell sheets without sutures.

Research Products

• [Journal Article] Comparison of the canine corneal epithelial cell sheets cultivated from limbal stem cells on canine amniotic membrane, atelocollagen gel, and temperature-responsive culture dish. (2015)
  • Author(s): Nam E, Fujita N, Morita M, Tsuzuki K, Lin HY, Chung CS, Nakagawa T, Nishimura R
  • Journal Title: Veterinary Ophthalmology Volume: 印刷中 Issue: 4 Pages: 317-325
  • DOI: 10.1111/vop.12241
  • Peer Reviewed / Open Access
• [Journal Article] Evaluation of ABCG2 and p63 expression in canine cornea and cultivated corneal epithelial cells (2014)
  • Author(s): Maresuke M., Fujita N., Takahashi A., Nam ER., Yui S., Chung CS., Kawahara N., Lin HY., Tsuzuki K., Nakagawa T., Nishimura R.
  • Journal Title: Veterinary Ophalmology Volume: (in press) Issue: 1 Pages: 59-68
  • DOI: 10.1111/vop.12147
  • Peer Reviewed / Open Access
• [Presentation] 犬輪部由来角膜上皮細胞の増殖能維持機構の検討 (2017)
  • Author(s): 森田希輔，藤田直己，佐伯亘平，林本考緒道，都築圭子，中川貴之，西村亮平
  • Organizer: 第16回日本再生医療学会総会
  • Place of Presentation: 仙台国際センター（宮城県仙台市）
  • Year and Date: 2017-03-06
• [Presentation] 増殖因子およびフィーダー細胞非存在下での培養による犬輪部由来角膜上皮細胞株の樹立とその性状解析 (2016)
  • Author(s): 森田希輔、藤田直己、林本考緒道、都築圭子、中川貴之、西村亮平
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 日本大学（神奈川県藤沢市）
  • Year and Date: 2016-09-06
• [Presentation] 猫角膜上皮細胞に対するネコリコンビナントＨＧＦの作用と点眼薬としての有用性 (2016)
  • Author(s): 阿部桃子、藤田直己、木ノ下千佳子、森田希輔、都築圭子、中川貴之、西村亮平
  • Organizer: 日本再生医療学会
  • Place of Presentation: 大阪国際会議場（大阪府・大阪市）
  • Year and Date: 2016-03-17
• [Presentation] 無フィーダーで長期継代維持された犬輪部由来角膜上皮細胞の性状解析 (2016)
  • Author(s): 森田希輔、藤田直己、阿部桃子、都築圭子、中川貴之、西村亮平
  • Organizer: 日本再生医療学会
  • Place of Presentation: 大阪国際会議場（大阪府・大阪市）
  • Year and Date: 2016-03-17
• [Presentation] Therapeutic effect of feline recombinant hepatocyte growth factor (fr-HGF) on corneal injury in dogs (2016)
  • Author(s): Maresuke Morita, Naoki Fujita, Keiko Tsuzuki, Chikako Kinoshita, Dai Nagakubo, Kanako Muta, Hsing-Yi Lin, Yuiko Tanaka, Koji Hayashimoto, Takayuki Nakagawa, Ryohei Nishimura
  • Organizer: 6th Annual Congress of Asian Society of Veterinary Surgery
  • Place of Presentation: 福岡国際会議場（福岡県福岡市）
  • Int'l Joint Research
• [Presentation] Regulation of soluble factors in mesenchymal stem cells cultured as feeder cells for cultivation of corneal epithelial cell sheets in dogs. (2014)
  • Author(s): Morita M, Fujita N, Nam ER, Abe M, Tsuzuki K, Nakagawa, T, Nishimura R.
  • Organizer: Asian Meeting of Veterinary Surgery
  • Place of Presentation: 大阪
  • Year and Date: 2014-12-06 – 2014-12-07
• [Remarks] 東京大学獣医外科学教室HP
  • URL: http://www.vm.a.u-tokyo.ac.jp/geka/