Research of the isolation of pancreatic stem / progenitor cells for the treatment of diabetes mellitus

以胰岛移植为基础的治疗1型糖尿病的成功，以及胰岛极度短缺，促使人们努力开发可再生的胰岛替代组织来源。由于缺乏特异性标志物和细胞培养方法的发展，在前瞻性分离胰腺干/祖细胞方面只做了很少的尝试。本研究通过细胞分选的方法从胚胎胰腺中分离出胰腺干/祖细胞。免疫组织化学研究表明，这两种不同的细胞表面标志分别在上皮细胞和周围间充质细胞中表达。RT-PCR和微阵列分析表明，在这些表面标志的组合下，胰腺干/祖细胞在特定的细胞群中得到了丰富。在体内分化过程中，这些细胞群具有自我更新和多能性的能力，包括形成胰岛素产生细胞。此外，移植物中的胰岛样细胞团和祖细胞的细胞内胰岛素C肽含量也与成人胰岛相似。此外，我们在体外获得了这些群体的胰岛素分泌细胞和胰高血糖素分泌细胞的分化。由于该策略是基于使用人类和啮齿动物共同的细胞表面标记进行细胞分选的，因此它可能会推动从人类胰腺干细胞/祖细胞驱动可移植的胰岛替代组织的策略。此外，我们成功地建立了特定的表面标记基因敲除小鼠，并开始分析这些小鼠的有趣的表型。

Success in islet transplantation-based therapies for type 1 diabetes mellitus and an extreme shortage of pancreatic islets has motivated efforts to develop renewable sources of islet-replacement tissue. Only a few attempts have been made at prospective isolation of pancreatic stem / progenitor cells, due to the lack of specific markers and the development of cell culture method. This study demonstrates the isolation of pancreatic stem / progenitor cells from the embryonic pancreas by cell sorting. Immunohistological study showed the two different cell surface markers are expressed in the epithelial cells and the surrounding mesenchymal cells, respectively. RT-PCR and microarray analysis demonstrated that pancreatic stem / progenitor cells are enriched in the specified cell population, with the combination of these surface markers. During in vivo differentiation, these cell populations have the ability for self-renewal and multipotency, including the formation of insulin-producing cells. Moreover, the islet-like cell clusters in the graft and the intracellular insulin C-peptide content in progenitor-derived cells were also observed to be comparable to that of the adult pancreatic islets. In addition, we obtained the differentiation of insulin-producing cells and glucagons-producing cells from these populations in vitro. Since the strategy is based on the cell sorting using cell surface markers common to human and rodents, it may promote strategies to drive transplantable islet-replacement tissues from human pancreatic stem / progenitor cells. Furthermore, we succeed in establishing the specified surface marker gene-knockout mice and start to analyze the interesting phenotype of these mice.