Graduate School Research Meeting Presentation-大学院歯学独立研究科第 75 回 大 学 院 研 究 科 発 表 会 プ ロ グ ラ ム
第8 8 回 中 間 発 表 会 プ ロ グ ラ ム
大学院学生等が、これまでの研究成果を発表します。
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場 所：実習館 2 階 総合歯科医学研究所セミナー室
日 時：2017 年 11 月 22 日（水）17 時 25 分 開会2017 年 11 月 22 日（水）17 時 25 分 開会
No.

17：25
1

［研究科発表］17：30～18：00
司会：増田教授2

演題名・発表者

発表区分・予定時間［中間発表］
18：00～18：30
司会：宇田川教授

審査委員

開会挨拶 高橋研究科長「外耳道のひずみで咀嚼回数をカウントする方法」
菅生 秀昭 ４年 顎口腔機能制御学講座 咀嚼機能解析学

主査：金 銅 教授副査：十 川 教授
岡 田 准教授「骨髄間質細胞層の頂点に位置するレプチン受容体陽性細胞は骨形成因子 主査：川 上 教授
副査：八 上 准教授
Runx2 を発現する」
荒 講師
楊 孟雨 3 年 硬組織疾患制御再建学講座 硬組織機能解析学

問合わせ先：本館 2 階

学事室（大学院）内線 2331

発表内容の要旨（課程博士）
Abstract of Presented Research (For the Doctoral Course)
学

籍番

号

Student ID No.
（ ふ り が な ）

氏

名
攻分

1508

入 学 年
Entrance Year

2015

年

Year

やん もんいう

Name in Full

専

ID♯G野Major Field

楊 孟雨
硬組織疾患制御再建学講座 硬組織機能解析学

主 指 導 教 員

宇田川 信之発 表 会 区 分

中間発表会

Chief Academic Advisor
Type of Meeting･

大学院研究科発表会･松本歯科大学学会

Midterm Meeting / Graduate school research meeting presentation /The Matsumoto Dental University Society

演題名 / Title of Presentation
骨髄間質細胞層の頂点に位置するレプチン受容体陽性細胞は骨形成因子 Runx２を発現する
Osteogenic Factor Runx2 Marks a Subset of Leptin Receptor-Positive Cells that Sit Atop the Bone
Marrow Stromal Cell Hierarchy

発表要旨 / Abstract
Purpose:Bone marrow mesenchymal stem and progenitor cells (BM-MSPCs) maintain homeostasis of bone tissue byproviding osteoblasts (Annu Rev Immunol 31:285-316, 2013). Although several markers have been identified forlabeling of MSPCs, these labeled cells still contain non-BM-MSPC populations. Studies have suggested thatMSPCs are observed as leptin receptor (LepR)-positive cells, whereas osteoblasts can be classified as positive for
Runx2, a master regulator for osteoblastogenesis (Dev Cell 29:340-9, 2014). On the other hand, it is known thatbone volume is increased by intermittent parathyroid hormone (1-34) [iPTH(1-34)] treatment due to accelerated
the bone formation (J Bone Miner Res 17:808,2002). However it is unclear whether the PTH anabolic effect isexerted by mediating the BM-MSPCs. In this study, we will try to understand the questions using by genetic mice.
Method:1. Analysis of LepR+ cells in the bone marrow cavity express Runx2.
We used Runx2-GFP reporter mice, in which GFP is driven by a bacterial artificial chromosome (BAC) of Runx2
locus (J Bone Miner Res 29:1960-1969, 2014). FITC-conjugated anti-GFP antibody was used to amplify theintensity of the GFP signal when imaging bone tissue section. After cryosections made, we analyzed the Runx2
expression in LepR+ cells.
2. Analysis of LepR+ cells contain Runx2-GFPlow and Runx2-GFP- sub-population.
We used Runx2-GFP reporter mice. The bone marrow was gently flushed in L-15 FACS buffer. BM was digested
with 0.1% collagenase IV, 0.2% Dispase and 20 U/ml DNase in HBSS. We did the cell sorting and flowcytometry.
3. Analysis of Stromal stem cell activity in BM in enriched in LepR+Runx2-GFPlow population.
We used LepR-cre mice, in which Leptin Receptor (LepR) positive cells have characteristics of BM-MSCs in vivoby fate mapping approach. (Dev Cell 29:340-9, 2014). We did CFU-F assay, Spheroid formation assay and RNA
isolation and quantitative real-time PCR.
4. Analysis of LepR+Runx-GFPlow cells differentiate into osteoblasts through multilayered cell formation inresponse to PTH-induced anabolic effects.
We generated LepR-Cre/Tomato/Runx2-GFP mice, which LepR+ cells-derived osteoblasts were observed asyellow cells. We used tamoxifen-administered iOsx/Tomato/Runx2-GFP mice, which mature osteoblasts express
both iOsx/Tomato and Runx2-GFP. We also generated iOsx/Tomato/Col1(2.3)-GFP mice, which Col1(2.3)-GFP
was a marker of mature osteoblasts. We made cryosections by these three kinds mice.
Results:
1. Runx2 is heterogeneously expressed in the LepR+ BM stromal cell population.
2. Stem cell activity is enriched in the Runx2-GFPlow sub-population of LepR+ cells.
3. LepR+Runx2-GFPlow cells differentiate into osteoblasts through multilayered cell formation in response to PTH
anabolic effects.
4. LepR+Runx2-GFPlow cell-derived multilayered cells differentiate into mature osteoblasts with increasing
expression of Osterix and type I collagen.
Conclusion:
Runx2 is weakly expressed in the LepR+ population without osteoblastic commitment, and theLepR+Runx2-GFPlow stromal cells sit atop the BM stromal hierarchy.

大学院歯学独立研究科 2005/01/12