Induction of Insulin-Producing Cells From Human Pancreatic Progenitor Cells
Induction of Insulin-Producing Cells From Human Pancreatic Progenitor Cells
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DOI：
10.1016/j.transproceed.2010.05.097
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发表时间：
2010-07-01
影响因子：
0.9
通讯作者：
Matsumoto, S.
中科院分区：
文献类型：
--
作者：
Noguchi, H.;Naziruddin, B.;Matsumoto, S.
导论.我们以前建立了一个没有遗传操作的小鼠胰腺干细胞系。在这项研究中，我们试图识别和分离人胰腺干/祖细胞。我们还测试了生长因子和胰腺和十二指肠同源框因子-1（PDX-1）和BETA2/NeuroD蛋白转导到人胰腺干/祖细胞中是否诱导胰岛素或胰腺相关基因表达。采用经埃德蒙顿方案改良的标准Ricordi技术，将来自脑死亡供体的人胰腺用于胰岛分离。在几种培养基中培养来自导管丰富群体的细胞，这些培养基基于为小鼠胰腺或人类胚胎干细胞设计的培养基。为了诱导细胞分化，将细胞与exendin-4、烟酰胺、角质形成细胞生长因子、PDX-1蛋白或BETA 2/NeuroD蛋白一起培养2周。无血清培养基中的细胞显示出与小鼠胰腺干细胞系相似的形态，而人类胚胎干细胞培养基中的细胞形成成纤维细胞样形态。在培养过程中，每种培养基中细胞的核/质比均降低。细胞在30天后停止分裂，表明它们已经进入衰老期。经诱导培养液处理的细胞可分化为胰岛素分泌细胞，并表达胰腺相关基因。从导管丰富的人口的细胞复制是有限的。几种生长因子和转导蛋白的诱导治疗可能为诱导可移植的胰岛素产生细胞提供一种潜在的新策略。
Introduction. We previously established a mouse pancreatic stem cell line without genetic manipulation. In this study, we sought to identify and isolate human pancreatic stem/progenitor cells. We also tested whether growth factors and protein transduction of pancreatic and duodenal homeobox factor-1 (PDX-1) and BETA2/NeuroD into human pancreatic stem/progenitor cells induced insulin or pancreas-related gene expressions.Materials and method. Human pancreata from brain-dead donors were used for islet isolation with the standard Ricordi technique modified by the Edmonton protocol. The cells from a duct-rich population were cultured in several media, based on those designed for mouse pancreatic or for human embryonic stem cells. To induce cell differentiation, cells were cultured for 2 weeks with exendin-4, nicotinamide, keratinocyte growth factor, PDX-1 protein, or BETA2/NeuroD protein.Results. The cells in serum-free media showed morphologies similar to a mouse pancreatic stem cell line, while the cells in the medium for human embryonic stem cells formed fibroblast-like morphologies. The nucleus/cytoplasm ratios of the cells in each culture medium decreased during the culture. The cells stopped dividing after 30 days, suggesting that they had entered senescence. The cells treated with induction medium differentiated into insulin-producing cells, expressing pancreas-related genes.Conclusion. Duplications of cells from a duct-rich population were limited. Induction therapy with several growth factors and transduction proteins might provide a potential new strategy for induction of transplantable insulin-producing cells.