Development of Optimal Cell Culture Method with Morphological Quality Assessment by DNA Damage

2016 Fiscal Year Final Research Report

• Project/Area Number: 26293422
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Dental engineering/Regenerative dentistry
• Research Institution: National Center for Geriatrics and Gerontology
• Principal Investigator: NAKASHIMA Misako 国立研究開発法人国立長寿医療研究センター, 幹細胞再生医療研究部, 部長 (20207773)
• Co-Investigator(Kenkyū-buntansha): 山本 徳則 名古屋大学, 医学(系)研究科(研究院), 准教授 (20182636) ; 村上 真史 国立研究開発法人国立長寿医療研究センター, その他部局等, その他 (30614531) ; 栗田 賢一 愛知学院大学, 歯学部, 教授 (40133483) ; 兼子 智 東京歯科大学, 歯学部, 講師 (40214457)
• Research Collaborator: IOHARA Koichiro ; AHMED Nermeen
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 歯髄幹細胞 / マルチガスインキュベータ / 微小重力培養 / 品質検査 / DNA損傷

Outline of Final Research Achievements

The present study aimed at development of the optimal culture method and quality control and assurance method for dental pulp stem cells (DPSCs). At first, the morphological quality assessment of genomic DNA damage was performed by Single-Cell Pulsed-Field Gel Electrophoresis (SCPFGE) as a quality control and assurance method. The enzymatic or physical detachment to get a single cell suspension, however, caused a nonspecific DNA damage. Next, the optimal hypoxic condition for DPSCs was investigated in a tri-gas incubator for stable culture. The proliferation rate, stem cell properties and trophic effect were the highest in 5% O2 culture. Furthermore, three-dimensional microgravity devise　were used. There was, however, little difference in the proliferation rate, cell stability and quality in microgravity compared with normal gravity in only one passage of culture. Further three passages of culture under microgravity resulted in little difference in these from normal gravity.

Free Research Field

医歯薬学