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Therapeutic targets against gastric cancer stem cells interacting with tumor microenvironment

Journal of Cancer Metastasis and Treatment 2018年第1期4卷 110-117页

作者： tomoyuki uchihara Takatsugu Ishimoto Atsuko Yonemura Hideo Baba Department of Gastroenterological Surgery Graduate School of Medical ScienceKumamoto UniversityKumamoto 860-8556Japan

Gastric cancer(GC)is a major cause of cancer-related deaths *** existe nee of cancer stem cells(CSCs)is known to be the main reas on for resista nee to an ticanceragents as well as for the development of distant *** CSCs themselves harbor self-renewal and differentiation abilities,the tumor microenvironment that surrounds CSCs provides secreted factors and supports angiogenesis and is thus responsible for the maintenance of their CSC *** current review provides information regarding the impact of the tumor microenvironment on gastric CSCs,which will support the development of novel therapeutic strategies for targeting gastric CSCs.

关键词： Gastric cancer stem cells stem cell markers tumor microenvironment gastric cancer treatment

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Utility of tricalcium phosphate and osteogenic matrix cell sheet constructs for bone defect reconstruction

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World Journal of Stem Cells 2015年第5期7卷 873-882页

作者： tomoyuki Ueha Manabu Akahane Takamasa Shimizu Yoshinobu uchihara Yusuke Morita Naoya Nitta Akira Kido Yusuke Inagaki Kenji Kawate Yasuhito Tanaka Department of Orthopedic Surgery Nara Medical University Department of Public Health Health Management and Policy Nara Medical University Faculty of Medicine Department of Biomedical Engineering Doshisha University Department of Arthroplasty and Regenerative Medicine Nara Medical University

AIM: To determine the effects of transplanting osteogenic matrix cell sheets and beta-tricalcium phosphate(TCP) constructs on bone formation in bone ***: Osteogenic matrix cell sheets were prepared from bone marrow stromal cells(BMSCs), and a porous TCP ceramic was used as a scaffold. Three experimental groups were prepared, comprised of TCP scaffolds(1) seeded with BMSCs;(2) wrapped with osteogenic matrix cell sheets; or(3) both. Constructs were implanted into a femoral defect model in rats and bone growth was evaluated by radiography, histology, biochemistry, and mechanical testing after 8 wk. RESULTS: In bone defects, constructs implanted with cell sheets showed callus formation with segmentalor continuous bone formation at 8 wk, in contrast to TCP seeded with BMSCs, which resulted in bone nonunion. Wrapping TCP constructs with osteogenic matrix cell sheets increased their osteogenic potential and resulting bone formation, compared with conventional bone tissue engineering TCP scaffolds seeded with BMSCs. The compressive stiffness(mean ± SD) values were 225.0 ± 95.7, 30.0 ± 11.5, and 26.3 ± 10.6 MPa for BMSC/TCP/Sheet constructs with continuous bone formation, BMSC/TCP/Sheet constructs with segmental bone formation, and BMSC/TCP constructs, respectively. The compressive stiffness of BMSC/TCP/Sheet constructs with continuous bone formation was significantly higher than those with segmental bone formation and BMSC/TCP ***: This technique is an improvement over current methods, such as TCP substitution, and is useful for hard tissue reconstruction and inducing earlier bone union in defects.

关键词： Bone marrow stromal cells Osteogenesis Bone regeneration Tissue engineering Calcium phosphate

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Role of LIGHT in the pathogenesis of joint destruction in rheumatoid arthritis

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World Journal of Experimental Medicine 2017年第2期7卷 49-57页

作者： Afsie Sabokbar Sara Afrough David J Mahoney Yoshinobu uchihara Catherine Swales Nicholas A Athanasou Nuffield Department of Orthopaedics Rheumatology and Musculoskeletal SciencesUniversity of OxfordNuffield Orthopaedic Centre

AIM To characterise the role of substitutes for receptoractivator nuclear factor kappa-B ligand(RANKL) in rheumatoid arthritis(RA) joint destruction. METHODS Synovial fluid(SF) macrophages isolated from the knee joint of RA patients were incubated with 25 ng/m L macrophage-colony stimulating factor(M-CSF) and 50 ng/mL LIGHT(lymphotoxin-like, exhibits inducible expression and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator, a receptor expressed by T lymphocytes) in the presence and absence of 25 ng/mL RANKL and 100 ng/mL osteoprotegerin(OPG) on glass coverslips and dentine slices. Osteoclastogenesis was assessed by the formation of multinucleated cells(MNCs) expressing tartrate-resistant acid phosphatase(TRAP) on coverslips and the extent of lacunar resorption pit formation on dentine slices. The concentration of LIGHT in RA and osteoarthritis(OA) synovial fluid was measured by an enzyme-linked immunosorbent assay(ELISA) and the expression of LIGHT in RA and OA synovium was determined by immunohistochemistry using an indirect immunoperoxidase technique. RESULTS In cultures of RA SF macrophages treated with LIGHT and M-CSF, there was significant formation of TRAP + MNCs on coverslips and extensive lacunar resorption pit formation on dentine slices. SF-macrophage-osteoclast differentiation was not inhibited by the addition of OPG, a decoy receptor for RANKL. Resorption pits were smaller and less confluent than in RANKL-treated cultures but the overall percentage area of the dentine slice resorbed was comparable in LIGHT-and RANKL-treated cultures. LIGHT significantly stimulated RANKL-induced lacunar resorption compared with RA SF macrophages treated with either RANKL or LIGHT alone. LIGHT was strongly expressed by synovial lining cells, subintimal macrophages and endothelial cells in RA synovium and the concentration of LIGHT was much higher in RA compared with OA *** LIGHT is highly expressed in RA synovium and SF, stimulate

关键词： Receptor-activator nuclear factor kappa-B ligand Osteoclast Rheumatoid arthritis Resorption LIGHT

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Use of Cryopreserved Osteogenic Matrix Cell Sheets for Bone Reconstruction

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Stem Cell Discovery 2016年第1期6卷 13-23页

作者： Tomohiko Kura Manabu Akahane Takamasa Shimizu Yoshinobu uchihara Yasuaki Tohma Yusuke Morita Munehisa Koizumi Kenji Kawate Yasuhito Tanaka Department of Orthopedic Surgery Nara Medical University School of Medicine Kashihara Japan Department of Public Health Health Management and Policy Nara Medical University School of Medicine Kashihara Japan Departmentof Biomedical Engineering Doshisha University Kyotanabe Japan Department of Artificial Joint and Regenerative Medicine Nara Medical University Kashihara Japan

Skeletal diseases, such as nonunion and osteonecrosis, are now treatable with tissue engineering techniques. Single cell sheets called osteogenic matrix cell sheets (OMCSs) grown from cultured bone marrow-derived mesenchymal stem cells show high osteogenic potential;however, long preparation times currently limit their clinical application. Here, we report a cryopreservation OMCS transplantation method that shortens OMCS preparation time. Cryopreserved rat OMCSs were prepared using slow- and rapid-freezing methods, thawed, and subsequently injected scaffold-free into subcutaneous sites. Rapid- and slow-frozen OMCSs were also transplanted directly to the femur bone at sites of injury. Slow-freezing resulted in higher cell viability than rapid freezing, yet all two cryopreservation methods yielded OMCSs that survived and formed bone tissue. In the rapid- and slow-freezing groups, cortical gaps were repaired and bone continuity was observed within 6 weeks of OMCS transplantation. Moreover, while no significant difference was found in osteocalcin expression between the three experimental groups, the biomechanical strength of femurs treated with slow-frozen OMCSs was significantly greater than those of non-transplant at 6 weeks post-injury. Collectively, these data suggest that slow-frozen OMCSs have superior osteogenic potential and are better suited to produce a mineralized matrix and repair sites of bone injury.

关键词： Bone Marrow Stromal Cell Cryopreservation Cell Sheet Injectable Bone Bone Reconstruction

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