Construction of vascular network-intrinsic peripheral nerve organoids using dental pulp stem cells by a novel cell bead method

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K21029
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0907:Oral science and related fields
• Research Institution: University of Tsukuba
• Principal Investigator: Takaoka Shohei 筑波大学, 附属病院, 医員 (30958567)
• Project Period (FY): 2022-08-31 – 2024-03-31
• Keywords: 間葉系幹細胞 / 歯髄幹細胞 / 神経誘導 / シングルセルRNAシーケンス / オルガノイド / 末梢神経

Outline of Final Research Achievements

We have successfully obtained neural lineage cells (NLCs) from human dental pulp stem cells. Single-cell RNA sequencing analysis showed that the NLCs consisted of neural progenitors, neurons, astrocytes, and a few non-neuronal lineage cells, and that the neural progenitors first produced neurons and then differentiated into astrocytes. In addition, a very small number of neural crest cells were also shown to be present.
NLC beads and endothelial cell beads were successfully cultured in perfusion in PDMS plates with straight lines connecting the holes to each other to promote axon elongation in the straight sections. Most of the axons formed myelin sheaths by Schwann cells, revealing the presence of a vascular network with a lumen.

Free Research Field

口腔再生医学

Academic Significance and Societal Importance of the Research Achievements

歯髄幹細胞を含む間葉系幹細胞の神経誘導は、神経関連たんぱくや遺伝子の発現によって特徴づけられることが多い。本研究は歯髄幹細胞の神経誘導の結果は単一細胞RNAシーケンス解析によってニューロンとアストロサイトを生み出す神経前駆細胞を特定した。これは中枢神経系疾患への細胞移植治療に大きな意味を持つ。
さらに末梢神経オルガノイド構築において、歯髄幹細胞の神経誘導の結果存在していた少数の神経堤細胞がシュワン細胞へ分化し髄鞘を形成したと考えられる。本研究で構築された血管網を内在した末梢神経オルガノイドは末梢神経をターゲットとした再生医療や創薬研究等に応用可能な実験モデルとなり得る。