Islet neogenesis-associated protein-related pentadecapeptide enhances the differentiation of islet-like clusters from human pancreatic duct cells
Islet neogenesis-associated protein-related pentadecapeptide enhances the differentiation of islet-like clusters from human pancreatic duct cells
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DOI：
10.1016/j.peptides.2009.09.003
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发表时间：
2009-12-01
期刊：
影响因子：
3
通讯作者：
Yang, Tao
中科院分区：
文献类型：
--
作者：
Li, Juan;Wang, Yun;Yang, Tao
将胰腺导管上皮细胞分化为β细胞被认为是增加移植胰岛数量的一种替代方法。在胰管细胞的体外分化方案中引入了关键因素，以促进胰岛β细胞的产生。胰岛新生相关蛋白(INGAP)是胰岛新生的启动子，INGAP的104-118肽序列可刺激动物的胰岛β细胞质量增加，也存在于人类参与胰岛新生的病理状态。为建立一种新的INGAP相关十五肽(INGAP-PP)诱导人胰管细胞分化的方法，采用烟酰胺、exendin-4、转化生长因子β(1)和INGAP-PP/杂乱多肽(SCHERBLED-P)四步法分离、纯化和扩增人胰管细胞。诱导分化后，INGAP-PP组的胰岛样簇(ILCs)的生成明显高于SCRAMED-P对照组。免疫荧光和逆转录聚合酶链式反应显示细胞角蛋白19、胰腺十二指肠同源盒-1、巢蛋白阳性表达，胰岛素和胰升糖素阴性。此外，在高糖刺激下，INGAP-PP组的ILCs分泌的胰岛素和C-肽水平高于SCHERFED-P组。结论：INGAP多肽可促进胰管细胞在体外分化为胰岛样簇。(C)2009 Elsevier Inc.保留所有权利。
The differentiation of pancreatic ductal epithelial cells into beta-cells has been considered as an alternative method for increasing the number of islets for transplantation. Critical factors have been introduced into the in vitro differentiation protocol for pancreatic duct cells in order to enhance the production of beta-cells. Islet neogenesis-associated protein (INGAP) is an initiator of islet neogenesis and the peptide sequence 104-118 of INGAP has been shown to stimulate an increase in beta-cell mass in animals and also found in human pathological states involving islet neogenesis. To establish a novel method for the differentiation of beta-cells from human pancreatic duct cells with INGAP-related pentadecapeptide (INGAP-PP), the pancreatic duct cells were isolated, purified and expanded in vitro and differentiated using a four-step protocol that included nicotinamide, exendin-4, transforming growth factor beta(1) and INGAP-PP/Scrambled peptide (Scrambled-P). The production of islet-like clusters (ILCs) in the INGAP-PP group was significantly higher than that in the Scrambled-P control group after differentiation from an equal number of duct cells. The duct cells showed positive staining and expression for cytokeratin 19, pancreatic duodenal homeobox-1, nestin, and were negative for insulin and glucagon, as detected by both immunofluorescence and RT-PCR Following differentiation the cells became insulin and glucagon positive. In addition, the ILCs from the INGAP-PP group secreted higher levels of insulin and C-peptide than the Scrambled-P group under a high glucose challenge. We conclude that INGAP pepticle enhances the in vitro differentiation of pancreatic duct cells into islet-like clusters. (C) 2009 Elsevier Inc. All rights reserved.