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Induction of insulin secreting cell and its application for transplantation

胰岛素分泌细胞的诱导及其在移植中的应用

基本信息

批准号：
14370346
负责人：
SUNAMURA Makoto
金额：
$ 9.73万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370346/
关键词：
pancreatic ductal epithelium insulin secreting cell pancreatic islet regeneration PDX-1 gene microarray analysis 膵管上皮細胞ラ島 PDX1 マイクロアレイ膵ラ島 VEGF angiopoietin 血管新生 PDX-1 膵再生

项目摘要

Islet transplantation has been expected to be effective therapy for insulin dependent DM (IDDM). However, this strategy is not established as a standard treatment for IDDM yet. The control of immuno-rejection is an obstacle for islet transplantation as same as the shortage of donors. The aim of this study is to develop the islet transplantation by using regeneration islet.1.The induction of insulin secreting cells from human pancreatic duct cell (HPDC)1)We established the HPDC cell line by the transfection of early expression gene of papilloma virus.2)The change of shape of this HPDC was observed after the stimulation of extra cellular matrix of tissue culture.3)We constructed the adenovirus vector for the transfection of PDX-1 gene. After transfection of PDX-1 gene into HPDC, the more expression of glucagon was detected than that of insulin. The cells made a mass looking like islet after in vitro culture. These HPDC became neoplastic tumor in vivo transplantation in mice.2.The analysis of genes expression using microarray and the detection of genes relating to islet development1)The mRNA was extracted from HPDC showing the insulin expression and used for DNA microarray study. The exspression of insulin was detected and genes showing the same pattern as insulin were selected.2)The collaborate group has established insulin secreting cell from mouse embryonic stem cell (ISSC). We harvested the cells from 5 stage of this development and analyzed the mRNA expression using microarray system. The analysis of changes in gene expression revealed that the expression of genes relating to insulin was increased step by step. Theses results indicate that the responsible gene would be detected by the further study.

胰岛移植有望成为治疗胰岛素依赖型糖尿病（IDDM）的有效方法。然而，该策略尚未被确立为IDDM的标准治疗。免疫排斥反应的控制和供体的短缺是胰岛移植的一大障碍。1.人胰管细胞胰岛素分泌细胞（HPDC）的诱导1）通过转染乳头瘤病毒早期表达基因建立HPDC细胞系2）观察HPDC在组织培养细胞外基质刺激下的形态变化3）观察HPDC的形态变化构建PDX-1基因的腺病毒载体。PDX-1基因转染HPDC后，胰高血糖素的表达明显高于胰岛素的表达。细胞体外培养后形成胰岛样团块。2.基因表达谱芯片分析及胰岛发育相关基因的检测1）从HPDC中提取胰岛素表达的mRNA，用于基因芯片研究。检测胰岛素基因的表达，筛选出与胰岛素基因表达模式相同的基因。2）本课题组从小鼠胚胎干细胞（ISSC）中建立了胰岛素分泌细胞。我们从这个发展的5个阶段收集细胞，并使用微阵列系统分析mRNA表达。对基因表达变化的分析表明，胰岛素相关基因的表达逐步增加。这些结果表明，通过进一步的研究，可以找到致病基因。