Development of Doxycycline-Dependent Inducible SV40LT-Immortalized Human Conjunctival Epithelial Cell Line in a three-dimensional culture and Its Characterization

Research Project

Project/Area Number	16H07026
Research Category	Grant-in-Aid for Research Activity Start-up
Allocation Type	Single-year Grants
Research Field	Ophthalmology
Research Institution	Ehime University
Principal Investigator	Mitani Arisa 愛媛大学, 医学部附属病院, 医員 (60648096)
Project Period (FY)	2016-08-26 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000) Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Keywords	不死化結膜上皮細胞株 / 三次元培養 / ムチン / 不死化結膜上皮細胞 / ドキシサイクリン誘導性 / 三次元培養システム / 細胞・組織
Outline of Final Research Achievements	Conjunctival epithelium is believed to play an important role in barrier formation and stabilization of tears on the ocular surface. However, due to the absence of suitable conjunctival epithelial cultured cells, detailed examination is difficult. In this study, a human conjunctival epithelial cell line capable of controlling the expression of SV40 large T antigen (SV40LT) gene was prepared according to the presence or absence of doxycycline. In addition, three - dimensional culture was carried out in which epithelial cells and parenchymal cells could be co - cultured in three dimensions, and multilayered conjunctival epithelial cells with markers characteristic of conjunctival epithelium could be confirmed. It is considered that it is possible to construct a culture conjunctival model that can maintain the characteristic differentiation potential of conjunctival epithelium in the long term in the future.