Optimization of regeneration technique for bone and cartilage derived from iPS cells

• Project/Area Number: 26293336
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Osaka University
• Principal Investigator: Yoshikawa Hideki 大阪大学, 医学(系)研究科(研究院), 理事・副学長 (60191558)
• Co-Investigator(Kenkyū-buntansha): 名井 陽 大阪大学, 医学部附属病院, 准教授 (10263261) ; 中田 研 大阪大学, 医学(系)研究科(研究院), 教授 (00283747) ; 中村 憲正 大阪大学, 国際医工情報センター, 招へい教授 (50273719) ; 梅澤 明弘 国立研究開発法人国立成育医療研究センター, その他部局等, その他 (70213486) ; 福田 寛二 近畿大学, 医学部, 教授 (50201744)
• Co-Investigator(Renkei-kenkyūsha): Hamaguchi Satoshi 大阪大学, 工学研究科, 教授 (60301826) ; Goto Naohisa 大阪大学, 微生物研究所, 講師 (60448157)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2016: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2015: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2014: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
• Keywords: 骨再生療法 / iPS細胞 / 骨芽細胞系細胞 / 骨芽細胞分化 / 間葉系前駆細胞 / 無血清培地 / 大型骨欠損 / 骨再生 / 軟骨 / 再生療法 / 培養誘導 / 骨

Outline of Final Research Achievements

Osteoblast lineage cells derived from mouse iPS cells were efficiently differentiated by only two-dimensional culture without inducing embryoid body formation. Directed osteoblast lineage cells were characterized in that they have not only high alkaline phosphatase activity, but also proliferation, migration ability and osteogenic potential, compared with mouse mesenchymal stem cells derived from bone marrow and calvarial cells. Moreover, the osteoblast lineage cells co-expressed in a part of Hox and Semaphorin genes.
We succeeded an efficient optimization for directing osteoblast differentiation from iPS cells in combination with administration of compounds and cytokines, in addition to culture conditions such as serum-free media, hypoxic culture and chemically modified dish surface.

Research Products

• [Journal Article] Oxygen ultra-fine bubbles water administration prevents bone loss of glucocorticoid-induced osteoporosis in mice by suppressing osteoclast differentiation (2017)
  • Author(s): Noguchi, T., Ebina, K., Hirao, M., Morimoto, T., Koizumi, K., Kitaguchi, K., Matsuoka, H., Iwahashi, T., Yoshikawa, H.
  • Journal Title: Osteoporosis International Volume: 28 Pages: 1063-1075
  • DOI: 10.1007/s00198-016-3830-1
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Comparison of 2 different formulations of artificial bone for a hybrid implant with a tissue-engineered construct derived from synovial mesenchymal stem cells (2017)
  • Author(s): Shimomura, K., Moriguchi, Y., Nansai, R., Fujie, H., Ando, W., Horibe, S., Hart, D.A., Gobbi, A., Yoshikawa, H., Nakamura N.
  • Journal Title: American Journal of Sports Medicine Volume: 45 Pages: 666-675
  • DOI: 10.1177/0363546516668835
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Vibration acceleration promotes bone formation in rodent models (2017)
  • Author(s): Uchida, R., Nakata, K., Kawano, F., Yonetani, Y., Ogasawara, I., Nakai, N., Mae, T., Matsuo, T., Tachibana, Y., Yokoi, H., Yoshikawa, H.
  • Journal Title: PLoS One. Volume: 12
  • DOI: 10.1371/journal.pone.0172614
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Synovial mesenchymal stem cells from osteo- or rheumatoid arthritis joints exhibit good potential for cartilage repair using a scaffold-free tissue engineering approach. (2016)
  • Author(s): Koizumi K, Ebina K, Hart DA, Hirao M, Noguchi T, Sugita N, Yasui Y, Chijimatsu R, Yoshikawa H, Nakamura N.
  • Journal Title: Osteoarthritis Cartilage Volume: Mar 10 Pages: 1413-1422
  • DOI: 10.1016/j.joca.2016.03.006
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Preparation of Scaffold-Free Tissue-Engineered Constructs Derived from Human Synovial Mesenchymal Stem Cells Under Low Oxygen Tension Enhances Their Chondrogenic Differentiation Capacity (2016)
  • Author(s): Yasui, Y., Chijimatsu, R., Hart, D.A., Koizumi, K., Sugita, N., Shimomura, K., Myoui, A.., Yoshikawa, H., Nakamura, N.
  • Journal Title: Tissue Eng Part A Volume: 22 Pages: 490-500
  • DOI: 10.1089/ten.tea.2015.0458
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Scaffold-free, stem cell-based cartilage repair (2016)
  • Author(s): Yasui, Y., Ando, W., Shimomura, K., Koizumi, K., Chijimatsu, R., Hamamoto, S., Kobayashi, M., Yoshikawa, H., Nakamura,N.
  • Journal Title: Journal of Clinical Orthopaedics and Trauma Volume: 7 Pages: 157-163
  • DOI: 10.1016/j.jcot.2016.06.002
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] MEK5 suppresses osteoblastic differentiation. (2015)
  • Author(s): Kaneshiro S, Otsuki D, Yoshida K, Yoshikawa H, Higuchi C.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 463 Pages: 241-247
  • DOI: 10.1016/j.bbrc.2015.05.035
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Progranulin plays crucial roles in preserving bone mass by inhibiting TNF-α-induced osteoclastogenesis and promoting osteoblastic differentiation in mice. (2015)
  • Author(s): Noguchi T, Ebina K, Hirao M, Kawase R, Ohama T, Yamashita S, Morimoto T, Koizumi K, Kitaguchi K, Matsuoka H, Kaneshiro S, Yoshikawa H.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 465(3) Pages: 638-43
  • DOI: 10.1016/j.bbrc.2015.08.077
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] IL-6 negatively regulates osteoblast differentiation through the SHP2/MEK2 and SHP2/Akt2 pathways in vitro (2014)
  • Author(s): Kaneshiro, S., Ebina, K., Shi, K., Higuchi, C., Hirao, M., Okamoto, M., Koizumi, K., Morimoto, T., Yoshikawa, H., Hashimoto, J.
  • Journal Title: Journal of Bone and Mineral Metabolism Volume: 32 Pages: 378-392
  • DOI: 10.1007/s00774-013-0514-1
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cyp26b1 within the growth plate regulates bone growth in juvenile mice (2014)
  • Author(s): Minegishi Y, Sakai Y, Yahara Y, Akiyama H, Yoshikawa H, Hosokawa K,Tsumaki N.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 454 Pages: 12-18
  • DOI: 10.1016/j.bbrc.2014.10.001
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cyclic compressive loading on 3D tissue of human synovial fibroblasts upregulates prostaglandin E2 via Cox-2 production without IL-1β and TNFα (2014)
  • Author(s): Shimomura, K., Kanamoto, T., Kita, K., Akamine, Y., Nakamura, N., Mae, T., Yoshikawa, H., Nakamura, N
  • Journal Title: Bone and Joint Research Volume: 3 Pages: 280-288
  • DOI: 10.1302/2046-3758.39.2000287
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Osteochondral Repair Using a Scaffold-Free Tissue-Engineered Construct Derived from Synovial Mesenchymal Stem Cells and a Hydroxyapatite-Based Artificial Bone (2014)
  • Author(s): 下村和範、中村憲正
  • Journal Title: Tissue Engineering Volume: - Pages: 2291-2304
  • DOI: 10.1089/ten.tea.2013.0414
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Osteochondral Tissue Engineering with Biphasic Scaffold: Current Strategies and Techniques (2014)
  • Author(s): 下村和範、中村憲正
  • Journal Title: Tissue Engineering Volume: - Pages: 468-476
  • DOI: 10.1089/ten.teb.2013.0543
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] バイオマテリアルによる骨再生：過去・現在・未来 (2016)
  • Author(s): 吉川秀樹
  • Organizer: 日本バイオマテリアル学会シンポジウム2016
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2016-11-21
  • Invited
• [Presentation] 新しい骨補填材に対する期待：臨床の立場から (2016)
  • Author(s): 吉川秀樹
  • Organizer: 日本バイオマテリアル学会シンポジウム2016
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2016-11-21
  • Invited
• [Presentation] 多能性幹細胞から分化誘導した骨芽細胞系細胞は、高い骨形成能を有し、SemaphorinおよびHox遺伝子を発現する (2016)
  • Author(s): 宮本 諭、吉田 清志、吉川 秀樹、名井 陽
  • Organizer: 第34回日本骨代謝学会学術集会
  • Place of Presentation: 大阪国際会議場
  • Year and Date: 2016-07-20
  • Invited
• [Presentation] 多能性幹細胞から分化誘導した骨芽細胞系細胞の特性 (2016)
  • Author(s): 宮本諭、濱本秀一、吉田清志、岡本美奈、奥崎大介、後藤直久、浜口智志、吉川秀樹、名井陽
  • Organizer: 日本再生医療学会
  • Place of Presentation: 大阪
  • Year and Date: 2016-03-17
• [Presentation] 人工骨による骨再生：過去・現在・未来 (2016)
  • Author(s): 吉川秀樹
  • Organizer: 第12回しまなみ骨・関節フォーラム
  • Place of Presentation: 松山
  • Invited
• [Presentation] Analysis of Amino Group Formation on Polystyrene Surfaces by Nitrogen-Hydrogen-Based Plasma Irradiation (2015)
  • Author(s): Gotoh, K., Itsuki, D., Michiro Isobe1, Sugimoto, S., Miyamoto, S., Myoui, A., Yoshikawa, H., Hamaguchi, S.
  • Organizer: American Vacuum Society 62nd International Syposium
  • Place of Presentation: San Jose, U. S. A.
  • Year and Date: 2015-10-18
  • Int'l Joint Research
• [Presentation] Optimization of culture conditions for directing osteogenesis differentiation of mouse induced pluripotent stem cells (2015)
  • Author(s): Satoshi Miyamoto, Akira Myoui, Daisuke Okuzaki, Naohisa Goto, Satoshi Hamaguchi, Hideki Yoshikawa
  • Organizer: International Society for Stem Cell Research, 2015 Annual Meeting
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 2015-06-24 – 2015-06-26
• [Presentation] Optimization of culture conditions for directing osteogenesis differentiation of mouse induced pluripotent stem cells (2015)
  • Author(s): Miyamoto, S.,Myoui, A., Okuzaki, D., Goto, N., Hamaguchi, S., Yoshikawa, H.
  • Organizer: International Society for Stem Cell Research 2015 Annual Meeting
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 2015-06-24
  • Int'l Joint Research
• [Presentation] Plasma Surface Modification of Cell Culture Plates (2014)
  • Author(s): Dai Itsuki, Satoshi Sugimoto, Satoshi Miyamoto, Akira Myoui, Hideki Yoshikawa, Satoshi Hamaguchi
  • Organizer: International symposium on non-equilibrium plasma and complex-system sciences
  • Place of Presentation: Bochum, Germany
  • Year and Date: 2014-09-30 – 2014-10-02
• [Presentation] 細胞培養プレートのプラズマ表面処理 (2014)
  • Author(s): 斉宮大,杉本敏司,宮本諭, 名井陽, 吉川秀樹, 浜口 智志
  • Organizer: 応用物理学会秋季学術講演会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-17 – 2014-09-20
• [Presentation] Modification of hydroxyapatite and polystyrene surface for cell culture by low-pressure plasmas (2014)
  • Author(s): Dai Itsuki, Tomoko Ito, Satoshi Sugimoto, Yu Moriguchi, Satoshi Miyamoto, Akira Myoui, Hideki Yoshikawa, Satoshi Hamaguchi
  • Organizer: International Conference on Plasma Medicine
  • Place of Presentation: Nara, Japan
  • Year and Date: 2014-05-18 – 2014-05-23
• [Presentation] Biological Effect of Plasma Processing on Ceramics Artificial Bone (2014)
  • Author(s): Akira Myoui
  • Organizer: International Conference on Plasma Medicine
  • Place of Presentation: Nara, Japan
  • Year and Date: 2014-05-18 – 2014-05-23
  • Invited