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谱，首先对参与皮瓣存活的表皮角质形成细胞、内皮细胞、真皮成纤维细胞和胎儿真皮成纤维细胞的细胞-细胞相互作用进行了测试。胎儿成纤维细胞（BALB-3 T3，克隆A3）通过白血病抑制因子（LIE '）相关受体和细胞内机制如STAT 3酪氨酸磷酸化受到良好调节的信号转导途径。将小鼠全长LIF cDNA永久转染入ALB-3 T3，克隆A3中，并将细胞传代。血管内皮生长因子（VEGF）以1、10、100 ng剂量依赖性地促进细胞生长。在无血清培养条件下，10 ng VEGF刺激1 ~ 30分钟和100 ng VEGF刺激1 ~ 60分钟，可诱导与LIF介导的细胞增殖密切相关的磷酸化Erkp 42/p44丝裂原相关蛋白（MAP）的表达， ...更多信息 Boyden双室分析使用人间充质干细胞（hMSC）（其是外胚层以及间充质谱系的关键角色）与其他成体细胞类型（例如表皮角质形成细胞、内皮细胞和真皮成纤维细胞）之间的相互作用进行。在上室培养hMSC，下室培养成体细胞16 h后，表皮角质形成细胞的迁移率显著高于内皮细胞和真皮成纤维细胞（分别为345.0± 61.50，36.2 ±14.45，63.8±24.81，p<0.01）。hMSC和表皮角质形成细胞的单层共培养通过超微结构分析证明了两种细胞类型之间的基底膜样蛋白的产生。碱性成纤维细胞生长因子（bFGF）刺激的hMSC与人工真皮模板一起加速皮肤缺损。这些结果可能表明，胎儿成纤维细胞，这是基因转染的LIF cDNA，是有用的细胞增殖和进一步的间充质干细胞可能是一个强大的工具，为再生医学。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)