Studies on Epithelial-mesenchymal Interaction in Gastrointestinal Tract.

胃肠道上皮间质相互作用的研究。

• 批准号: 07457134
• 负责人: KAWANO Sunao
• 金额: $ 4.8万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457134/
• 关键词: gastric mucosa; proliferation; differentiation; 3-dimensional culture; epithelial-mesenchymal; interaction; growth factor; 3次元組織培養; 線維芽細胞増殖分化促進因子

In this study, we investigated epithelial-mesenchymal interaction between gastrointestinal (GI) epithelial cells and fibroblasts using a three dimensional culture system. All the fibroblast cell line examined (NIH 3T3, Sweiss 3T3, MRC-5, NRK) produced an activity that promote growth/survival of non-transformed epithelial cells from the Gl tract (RGM-1, IEC-6, and primary fetal rabbit gastic epithelial cells). Non-trasformed epithelial cell lines and primary gastric epithelial cells secreted an activity that promote growth/survival and morphological transformation of the fibroblasts, whereas this activity was lost in transformed epithelial cell lines (MKN-28,45, DLD-1, HepG2, COS-1).RGM-1, a non-transformed gastric epithelial cell line, produced an activity (s) that promote growth/survival and morphological transformation of fibroblasts and Ito cell and stimulates neurite outgrowth of PC-12 neuronal cell. To purify this activity (s), we collected 30 L of serum-free conditioned medium from large scale culture of RGM-1 cell. The activity was purified by a combination of heparin affinity, ion exchange, gel filtration, and reverse phase chromatography using a FPLC system. The purified activity has a molecular weight of 30 kDa and its pertial amino acid sequence was novel.

在这项研究中，我们研究了胃肠（GI）上皮细胞和成纤维细胞之间的上皮间质相互作用，使用三维培养系统。所有检查的成纤维细胞系（NIH 3 T3、Sweiss 3 T3、MRC-5、NRK）产生促进来自胃肠道的非转化上皮细胞（RGM-1、IEC-6和原代胎兔胃上皮细胞）的生长/存活的活性。未转化的上皮细胞系和原代胃上皮细胞分泌促进成纤维细胞生长/存活和形态转化的活性，而这种活性在转化的上皮细胞系中丧失（MKN-28，45，DLD-1，HepG 2，COS-1）.RGM-1，一种非转化的胃上皮细胞系，产生促进成纤维细胞和Ito细胞的生长/存活和形态转化并刺激PC-12神经元细胞的神经突生长的活性。为了纯化该活性，我们从RGM-1细胞的大规模培养物中收集30 μ L无血清条件培养基。使用FPLC系统通过肝素亲和、离子交换、凝胶过滤和反相色谱的组合纯化活性。纯化的活性分子量为30 kDa，其部分氨基酸序列是新的。

项目成果

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