Enrichment and functional analysis of human deciduous dental pulp stem cells for tooth regeneration

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K10165
• 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: Kagoshima University
• Principal Investigator: Inada Emi 鹿児島大学, 医歯学域鹿児島大学病院, 講師 (30448568)
• Co-Investigator(Kenkyū-buntansha): 佐藤 正宏 国立研究開発法人国立成育医療研究センター, ゲノム医療研究部, 共同研究員 (30287099) ; 齊藤 一誠 朝日大学, 歯学部, 教授 (90404540) ; 野口 洋文 琉球大学, 医学(系)研究科(研究院), 教授 (50378733)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 遺伝子工学的手法 / アルカリホスファターゼ / ヒト乳歯歯髄細胞

Outline of Final Research Achievements

Human deciduous dental pulp cells (HDDPC) were prepared as single cells by a limiting dilution method, and the expression of stem cell-specific genes was examined by RT-PCR. The results showed that the expression patterns of stem cell-specific genes such as alkaline phosphatase (ALP) differed among the cell clones tested. Furthermore, when we performed repeated gene transfer of Yamanaka’s factors into HDDPC to generate stem cell-like cells, just prior to iPS cell formation, there observed the presence of ALP-positive cells after the fourth gene transfer. This suggests that HDDPC can be induced to generate stem cell-like cells.
On the other hand, we successfully obtained a cell mass with 3-D structure comprised by highly differentiated cells, when naive iPS cells are induced to form embryoid bodies (EBs) and these EBs were subjected to organ culture system which had been developed in-house.

Free Research Field

小児歯科

Academic Significance and Societal Importance of the Research Achievements

再生医療では、自己再生能と分化能を有する幹細胞が重要な役割を果たす。脱落乳歯や智歯、矯正治療のために便宜的に抜歯される永久歯から採取される歯髄細胞は、組織再生のための有望な細胞ソースと考えられている。本研究により、ALP(+)HDDPCが多能性幹細胞の特性を有し、それが歯形成に関与する能力が歯組織再構成実験から証明されれば、歯学分野、その他の基礎研究分野への波及効果は大いに期待できる。