Characterization of neuron-like cells differentiated from human dental pulp stem cells

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K10172
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57070:Developmental dentistry-related
• Research Institution: Nihon University
• Principal Investigator: SHIRAKAWA Tetsuo 日本大学, 歯学部, 特任教授 (00187527)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: ヒト歯髄 / エピジェネティクス / 間葉系幹細胞 / 神経細胞 / レット症候群

Outline of Final Research Achievements

Mesenchymal stem cells (MSCs) derived from the dental pulp of a girl with Rett syndrome and healthy girls were cultured in a stem cell-specific medium. Among the 88 cell surface antigens examined, only CD69 was strongly expressed in MSCs derived from the Rett girl. The expression of CD69 mRNA increased in a concentration-dependent manner upon TNF-α stimulation. Subsequently, hTERT gene transfection was attempted to immortalize the MSCs derived from the Rett girl as a necessary step for their differentiation into neurons. While the transfection of G418-resistant hTERT gene using Lipofectamine into MSCs from the healthy girls was successfully achieved, the same process was not completed in the MSCs from the Rett girl because of a serious decline of cell activity during the hTERT gene transfection. As a result, differentiation of MSCs derived from the Rett girl into neurons was not achieved.

Free Research Field

小児歯科学

Academic Significance and Societal Importance of the Research Achievements

レット症候群女児歯髄由来の間葉系幹細胞（MSCs）について、健常女児歯髄由来のMSCsと比較して明らかに増殖能が低くまた老化が早いことに加えて、Lipofectamineによる遺伝子導入処理に脆弱であることが判明した。レット症候群女児由来MSCsに強発現していたCD69はリンパ球の早期活性化マーカーとして知られているが、レット症候群の症状や遺伝子導入処理での細胞脆弱性との関連は不明である。CD69については近年、脳常在性の制御性T細胞での発現が報告されており、制御性T細胞が脳内での過剰な炎症反応を抑制していることが示唆されることから、レット症候群の病態解明における標的分子の１つと考えられる。