Induction of insulin secreting cell and its application for transplantation

胰岛移植有望成为胰岛素依赖型糖尿病（IDDM）的有效治疗方法。然而，这一策略尚未被确定为IDDM的标准治疗方法。与供体短缺一样，免疫排斥反应的控制也是胰岛移植的一大障碍。本研究的目的是利用再生胰岛进行胰岛移植。人胰管细胞（HPDC）胰岛素分泌细胞的诱导1)通过转染乳头瘤病毒早期表达基因建立了人胰管细胞（HPDC）细胞系。2)组织培养细胞外基质刺激后，观察到该HPDC的形状变化。3)构建了转染PDX-1基因的腺病毒载体。PDX-1基因转染后，胰高血糖素的表达高于胰岛素的表达。细胞在离体培养后形成一个类似胰岛的团块。这些HPDC在小鼠体内移植后变成了肿瘤。基因芯片表达分析及胰岛发育相关基因检测1)从人乳头状瘤细胞中提取胰岛素表达mRNA，用于DNA芯片研究。检测胰岛素的表达，选择与胰岛素模式相同的基因。2)课题组建立了小鼠胚胎干细胞胰岛素分泌细胞（ISSC）。我们收集了5个发育阶段的细胞，并使用微阵列系统分析了mRNA的表达。基因表达变化分析显示胰岛素相关基因的表达逐渐增加。这些结果表明，在进一步的研究中，可能会发现相关的基因。

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.