Artificial endocrine organ composed of cultured human skin cells.

由培养的人体皮肤细胞组成的人工内分泌器官。

• 批准号: 07671334
• 负责人: INOKUCHI Sadaki
• 金额: $ 1.47万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-07671334/
• 关键词: human keratinocytes; diabetes; insulin; gene therapy; immunodeficient mice; cell transplantation; adenovirus vector; SCIDマウス

We proposed to produce artificial endocrine organ composed ofcultured human skin cells into which a human hormone gene was introduced.Human preproinsulin C-DNA driven under the methallothionein, MTT,gene promoter was cloned into a adenovirus vector. After infection of the virus, rAdMTTins, cultured human keratinocytes produced a significant amount of proinsulin in vitro. When the virus infected cells were transplanted into immunodeficient mice, balb/c nu/scid, decreased blood sugar concentration was observed for 3 days. During the same period, plasma proinsulin concentration of the mice was significantly elevated.These results indicate that the virus infected keratinocytes transiently secrete bioactive proinsulin into blood stream of the immunodeficient mice. For stable expression of the introduced gene, we are improving a retrovirus vector for gene transfar into human keratinocytes.

将人激素基因导入体外培养的人皮肤细胞，构建人工内分泌器官，在甲硫蛋白基因启动子的驱动下，将人胰岛素原C-DNA克隆入腺病毒载体。培养的人角质形成细胞感染rAdMTTins病毒后，在体外产生大量的胰岛素原。将病毒感染的细胞移植到免疫缺陷小鼠Balb/c nu/SCID中，观察到血糖浓度下降3天。在同一时期，小鼠的血浆胰岛素原浓度显著升高。这些结果表明，感染病毒的角质形成细胞瞬时将生物活性胰岛素原分泌到免疫缺陷小鼠的血流中。为了稳定表达导入的基因，我们正在改进一种逆转录病毒载体，用于将基因导入人角质形成细胞。

项目成果

Inokuchi S et al.: "Effects of fibroblasts of different origin on long term maintenance of xenotransplanted human epidermal keratinocytes in immunodeficient mice." Cell & Tissue Res.281. 223-229 (1995)

Inokuchi S 等人：“不同来源的成纤维细胞对免疫缺陷小鼠异种移植的人表皮角质形成细胞长期维持的影响”。

Yajima H et al.: "Construction and characterization of a recombinant adenovirus vector carrying the human preproinsulin gene under the control of the methallothionein gene promoter." BBRC. 229. 778-787 (1996)

Yajima H 等人：“在甲硫氨酸基因启动子控制下携带人前胰岛素原基因的重组腺病毒载体的构建和表征。”

Yajima H et al.: "Construction and Characterization of a Recombinant Adenovirus Vector Carrying the human Preproinsulin Gene under the Control of the Methallothlonein Gene Promoter." Biochemical and Biophysical Research Communications. 229. 778-787 (1996)

Yajima H 等人：“在甲硫磷基因启动子的控制下携带人前胰岛素原基因的重组腺病毒载体的构建和表征”。

Yajima H et al..: "Construction and Characterization of a Recoom blnant Adenovirus Vect or Carrying the human Preproinsulin Gene under the Control of the Methallothionein Gene Promoter." Biochemical and Biophysical Research Communications. 229. 778-787 (1

Yajima H等人：“在甲硫氨酸基因启动子的控制下携带人类前胰岛素原基因的重组腺病毒载体的构建和表征。”

Inokuchi S et al.: "Inokuchi S et al. Effects of fibroblasts of different origin on long term main tenance of xenotransplanted human epidermal keratinocytes in immunodeficient mice." Cell & Tissue Research. 281. 223-229 (1995)

Inokuchi S 等人：“Inokuchi S 等人。不同来源的成纤维细胞对免疫缺陷小鼠异种移植的人表皮角质形成细胞的长期维持的影响。”