The receptor tyrosine kinase expression in the flap and the gene therapy

受体酪氨酸激酶在皮瓣中的表达及基因治疗

• 批准号: 14571715
• 负责人: TANAKA Katsumi
• 金额: $ 1.86万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571715/
• 关键词: flap viavility; gene therapy; signal transduction; Erk-phosphorylation; Leulemia Inhibitory Factor, LIF; mesenchyal stem cell; Boyden Chamber; 人工真皮; 情報伝達機構; 情報伝達; ヒト骨髄間葉系幹細胞; ヌードラット; 移植再生; 塩基性線維芽細胞増殖因子; 骨形成因子

In order to investigate the receptor tyrosine kinase profiles, each component involving in the flap viability, such as epidermal keratinocyte, endothelial cell, dermal fibroblast and fetal dermal fibroblast was at first tested on the cell-to-cell interaction. The fetal fibroblast (BALB-3T3, clone A3) is well regulated signal-transduction pathway via leukemia inhibitory factor (LIE') associated receptor and intracellular mechanism such as STAT3 tyrosine phosphorylatiorn. The permanently transfected mouse full-length LIF cDNA into the ALB-3T3, clone A3 and the cell is passaged. The vascular endothelial growth facto (VEGF) significantly increased the cell growth in the doseincrease 1,10,100ng, dependent-manner. The phosphorylated-Erk p42/p44 Mitogen-Associated Protein (MAP), which is highly related to the proliferation of LIF-mediated cell, is induced from 1 to 30 minutes after 10ng VEGF and from l to 60 minutes after 100ng VEGF stimulation in the serum-free medium condition.The modified … More Boyden dual chamber analyses were performed with interaction between the human mesenchymal stem cell (hMSC), which is a key role player of the ectodermal as well as mesenchymal lineage, and other, adult cell types such as epidermal keratinocyte, endothelial cell and dermal fibroblast. In 16-hour incubation with the hMSC in the top chamber and adult cells in the lower chamber demonstrated the significant cell migration in epidermal keratinocyte compared to endothelial cell and dermal fibroblast (345.0±61.50,36.2±14.45, 63.8±24.81, respectively, p<0.01). The monolayer co-culture of hMSC and epidermal keratinocyte demonstrated basal membrane-like protein production between the two cell types by ultrastructural analyses. The hMSC stimulated by basic fibroblast growth factor (bFGF) accelerated the skin defect together with artificial dermis template. These results may indicate the fetal fibroblast, which is gene-transfected of LIF cDNA, is useful in cell proliferation and further the mesenchymal stem cell may be a powerful tool for regenerative medicine. Less

为了研究受体酪氨酸激酶的特性，首先检测了与皮瓣存活有关的各个成分，如表皮角质形成细胞、内皮细胞、真皮成纤维细胞和胎儿真皮成纤维细胞之间的相互作用。胎儿成纤维细胞(BALB-3T3，克隆A3)是通过白血病抑制因子(Lie‘)相关受体和STAT3酪氨酸磷酸化等细胞内机制调控的信号转导通路。将小鼠LIF全长c DNA永久性地转入ALB-3T3，克隆A3，细胞传代。血管内皮细胞生长因子(VEGF)以剂量增加1、10、100 ng的方式显著促进细胞生长。在无血清培养条件下，与激光诱导因子介导的细胞增殖密切相关的磷酸化ERK p42/p44丝裂原相关蛋白(MAP)在10 ng血管内皮生长因子刺激后1~30分钟和L刺激100 ng血管内皮细胞生长因子刺激后60分钟被诱导。改良的…更多的Boyden双室分析是在人类间充质干细胞(HMSC)与其他成人细胞类型(如表皮角质形成细胞、内皮细胞和真皮成纤维细胞)之间的相互作用下进行的，hMSC是外胚层和间质谱系的关键角色。与上室和下室成体细胞共同孵育16h，表皮角质形成细胞较内皮细胞和真皮成纤维细胞迁移明显(分别为345.0±61.5 0，36.2±14.45，63.8±2 4.81，P<0.0 1)。超微结构分析显示，hMSC与表皮角质形成细胞单层共培养可见两种细胞间产生基底膜样蛋白。碱性成纤维细胞生长因子(BFGF)刺激的hMSC与人工真皮模板共同促进皮肤缺损的修复。这些结果表明，转LIF基因的胎儿成纤维细胞可用于细胞增殖，间充质干细胞可能成为再生医学的有力工具。较少

项目成果

Akita S, Fukui M, Nakagawa H, Fujii T, Akino K: "Cranial bone defect healing is accelerated by mesenchymal stem cell by co-administration of bone morphogenetic protein-2 and basic fibroblast growth factor"Wound Repair Regen. 12. 254-261 (2004)

Akita S、Fukui M、Nakakawa H、Fujii T、Akino K：“间充质干细胞通过骨形态发生蛋白 2 和碱性成纤维细胞生长因子的共同施用加速颅骨缺损愈合”伤口修复再生。

Daian T, Ohtsuru A, Rogounovitch T, Ishihara H, Hirano A, Akiyama-Uchida Y, Saenko V, Fujii T, Yamashita S.: "Overexpression of insulin-like growth factor-1 (IGF-I) receptor and the invasiveness of cultured keloidfibroblasts."J invest Dermatol.. 120. 956-

Daian T、Ohtsuru A、Rogounovitch T、Ishihara H、Hirano A、Akiyama-Uchida Y、Saenko V、Fujii T、Yamashita S.：“胰岛素样生长因子-1 (IGF-I) 受体的过度表达及其侵袭性

Fukui M, Akita S, Akino K.: "Ectopic bone, formation facilitated by human mesenchymal stem cells and osteogenic cytokines via nutrient vessel injection in a nude rat model."Wound Rep Reg.. (in press).

Fukui M、Akita S、Akino K.：“通过在裸鼠模型中注射营养血管，人类间充质干细胞和成骨细胞因子促进异位骨的形成。”Wound Rep Reg..（出版中）。

田中克己: "形成外科におけるマイクロサージェリーの現況"日本生殖外科学会雑誌. 15:1. 26-34 (2002)

Katsumi Tanaka：“整形外科显微外科的现状”日本生殖外科学会杂志15：1（2002）。

Akino K, Mineta T, Fukui M, Fujii T, Akita S.: "Bone morphogenetic protein-2 regulates proliferation of human mesenchymal stem cells."Wound Rep Reg.. 11. 354-360 (2003)

Akino K、Mineta T、Fukui M、Fujii T、Akita S.：“骨形态发生蛋白 2 调节人类间充质干细胞的增殖。”Wound Rep Reg.. 11. 354-360 (2003)