Development of islet regeneration therapy by stem cell biology

干细胞生物学开发胰岛再生疗法

• 批准号: 13854020
• 负责人: SUMI Shoichiro
• 金额: $ 44.84万
• 依托单位: Institute for Frontier Medical Sciences, Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (S)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13854020/
• 关键词: embryonic stem cell; bio-artificial pancreas; immunoisolation; Langerhans islet (pancreatic islet); subcutaneous transplantation; neovascularization; diabetes mellitus; induction of differentiation; 幹細胞; ES細胞; 膵島様細胞塊; 塩基性線維芽細胞増殖因子; インスリン産生細胞

On the induction of differentiation toward insulin-producing cells, we successfully induced islet-like cell clusters (ICCs) form mouse embryonic stem cells by our own method. We showed that the ICCs contain electron microscopy-proved insulin granules, express pancrease-specific genes and reduce blood glucose levels of diabetic mice by transplantation.On the development of bio-artificial pancreas of possible clinical subcutaneous transplantation, we established rod-type device to normalize hyperglycemia of diabetic mice using rat islets and porcine pancreatic endocrine cells as bioreactor and anti-complement polymer as immunoisolation membrane when transplanted into prevascularized subcutaneous site. In addition, we developed a novel method to make macroencapsulated islets using polyvinyl alcohol hydrogel as semipermeable membrane by freezing/thawing method. This method is much easier in sealing islets in the gel and suitable for making larger devices in comparison to our and reported previous methods.On the methods to induce neovascularization for islet transplantation, we showed that gelatin microbeads impregnated with basic fibroblast growth factor in cooperation with collagen sponge has an excellent ability of vascular induction for islet transplantation in subcutaneous site. We further studied improved methods in ischemic rat skin flap models and newly developed rat leg model of prolonged ischemia.From these results, we consider that the system of subcutaneously implantable bio-artificial pancreas has been established at least for mouse diabetic models. On this basis, we are now ready to develop larger bio-artificial pancreas devices for preclinical examination in diabetic larger animals including dogs.

在向胰岛素分泌细胞的诱导分化方面，我们成功地用我们自己的方法从小鼠胚胎干细胞中诱导出胰岛样细胞团（ICCs）。我们发现ICCs含有电镜证实的胰岛素颗粒，表达胰腺特异性基因，并通过移植降低糖尿病小鼠的血糖水平。关于开发可能临床皮下移植的生物人工胰腺，我们以大鼠胰岛和猪胰腺内分泌细胞为生物反应器，建立了一种用于糖尿病小鼠血糖正常化的杆状装置，当移植到预先血管化的皮下部位时，补体聚合物作为免疫隔离膜。此外，我们还开发了一种以聚乙烯醇水凝胶为半透膜，采用冻融法制备大囊化胰岛的新方法。在胰岛移植的血管诱导方法上，我们发现明胶微球结合胶原海绵具有良好的血管诱导能力，可用于皮下胰岛移植。我们进一步研究了改进的大鼠皮瓣缺血模型和新开发的大鼠腿部长时间缺血模型，认为皮下植入式生物人工胰腺系统至少在小鼠糖尿病模型上已经建立。在此基础上，我们现在准备开发更大的生物人工胰腺装置，用于包括狗在内的糖尿病大型动物的临床前检查。

项目成果

Wang WJ, Gu Y, Tabata Y, Miyamoto M, Hori H, Nagata N, Toma M, Balamurugan AN, Kawakami Y, Nozawa M, Inoue K.: "Reversal of diabetes in mice by xenotransplantation of bioartificial pancreas in a prevascularized subcutaneous site."Transplantation. 73. 122-

Wang WJ、Gu Y、Tabata Y、Miyamoto M、Hori H、Nagata N、Toma M、Balamurugan AN、Kawakami Y、Nozawa M、Inoue K.：“通过在血管前皮下部位进行生物人工胰腺异种移植来逆转小鼠糖尿病

Xu B.Y.et al.: "The innuence of anticomplement synthetic sulfonic polymers on function of pancreatic islets : an in vitro study"Cell Transplant. 10. 413-417 (2001)

Xu B.Y.等人：“抗补体合成磺酸聚合物对胰岛功能的影响：体外研究”细胞移植。

Wang W.J.et al.: "Reversal of diabetes in mice by xenotransplantation of bioartificial pancreas in a prevascularized subcutaneous site"Transplantation. 73. 122-129 (2002)

Wang W.J.等人：“通过在预血管化的皮下部位进行生物人工胰腺异种移植来逆转小鼠糖尿病”移植。

Qi MG, Gu Y, Sakata N, Kim DH, Shirouzu Y, Yamamoto C, Hiura A, Sumi S, Inoue K.: "PVA hydrogel sheet macroencapsulation for the bioartificial pancreas"Biomaterials. (発表予定).

Qi MG、Gu Y、Sakata N、Kim DH、Shirouzu Y、Yamamoto C、Hiura A、Sumi S、Inoue K.：“生物人工胰腺的 PVA 水凝胶片宏观封装”（待提交）。

Kawakami Y. et al.: "Modified subcutaneous tissue with neovasclarization as the site of pancreatic islet translantation"Cell Transplant. 9. 729-732 (2001)

Kawakami Y.等人：“用新血管化修饰的皮下组织作为胰岛移植的部位”细胞移植。