| Project/Area Number |
16H05452
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
Kubo Toshikazu 京都府立医科大学, 医学(系)研究科(研究院), 特任教授 (20178031)
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| Co-Investigator(Kenkyū-buntansha) |
新井 祐志 京都府立医科大学, 医学(系)研究科(研究院), 准教授 (50347449)
中川 周士 京都府立医科大学, 医学(系)研究科(研究院), 助教 (30643382)
井上 裕章 京都府立医科大学, 医学(系)研究科(研究院), 助教 (60457968)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000)
Fiscal Year 2018: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2017: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2016: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
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| Keywords | 軟骨 / 変形性関節症 / 再生医療 / 幹細胞 / 物理刺激 / 整形外科学 / 軟骨分化 / 再生医学 / 硝子軟骨 / iPS細胞 |
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| Outline of Final Research Achievements |
We aimed to determine whether cartilaginous tissue can be produced from iPSCs under hypoxia and to evaluate the effects on the cellular metabolism and purity of the tissue. Human iPSCs were cultured for cartilage differentiation in monolayers under normoxia or hypoxia. We evaluated chondrocyte differentiation by real-time RT-PCR and FACS. Then, iPSCs were cultured for cartilage differentiation in 3D culture under normoxia or hypoxia. We evaluated the tissues on days 28 and 56 through histological analyses. Hypoxia suppressed the expression of immature mesodermal markers and promoted the expression of the chondrogenic markers. SOX9-positive cells were increased by culture under hypoxia. Percentages of safranin O-positive and type 2 collagen-positive tissues were increased under hypoxia. Hypoxia not only led to enhanced cartilage matrix production but also improved cell purity. By applying this method, highly pure cartilaginous-like tissues may be produced more rapidly and conveniently.
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| Academic Significance and Societal Importance of the Research Achievements |
近年，幹細胞から培養した軟骨組織移植を含めた再生医療が新たな治療法として有力視されている．特にiPSCは，細胞ソースとして優れている．しかしiPSCを用いた再生医療にも費用，培養期間，組織純度といった問題点がある．iPSCから軟骨組織を作製する方法はすでに確立されているが純度や組織作製にかかる期間が問題となっている。今回われわれの低酸素刺激を加えることで軟骨組織をより早期に、高純度で作製できる可能性があると考えた。本研究により今後iPSCを用いた軟骨組織移植が臨床応用するに当たり、物理刺激を加えることで、早期作製、高純度、低コストでの組織作製が可能になる可能性を見いだした点で意義深いと考える。
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