Identification of functional transcription factor in mouse gingival junctional epithelial cell using a bioengineered tooth system

Research Project

Project/Area Number	17K17359
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Periodontology
Research Institution	Showa University
Principal Investigator	Himuro Sara 昭和大学, 歯学部, 助教 (90736513)
Project Period (FY)	2017-04-01 – 2022-03-31
Project Status	Completed (Fiscal Year 2021)
Budget Amount *help	¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000) Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000) Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords	接合上皮
Outline of Final Research Achievements	We have succeeded in isolating JE cells expressing green fluorescent protein (GFP) by using a bioengineered tooth technique in mice. The gene expressions of GFP-positive JE cells, isolated from around the erupted bioengineered teeth using flow cytometry, were analyzed by RNA sequencing. GFP-positive cells derived from the bioengineered tooth germs showed similar gene expression patterns to primary JE cells. The isolated GFP-positive JE cells were immortalized by transducing the simian virus 40 large T antigen using lentiviral vectors. The established GFP-positive JE cells maintained proliferative activity for more than 20 passages, and did not show cellular senescence as demonstrated by β-galactosidase assay. These cells also expressed similar gene expression patterns to primary JE cells. The established cell lines may prove useful for future investigation of JE characteristics in vitro.
Academic Significance and Societal Importance of the Research Achievements	接合上皮は歯周病の起始点となる重要な組織でありながら、今まで、接合上皮細胞株が樹立されていなかった。今回、天然の接合上皮株と遺伝子発現パターンが酷似した不死化GFP陽性接合上皮株を樹立したことにより、in vitroの接合上皮細胞の研究が進むことが予想され、歯周病発症の機序の解明に繋がると考える。