Abstract:

背景：目前由于胰岛来源匮乏，使得胰岛细胞移植治疗糖尿病无法满足临床需求，故体外将胰腺干细胞诱导分化为胰岛成为研究焦点。
目的：于体外将小鼠胰腺干细胞诱导成胰岛样细胞团并对其进行相关检测，探寻一种胰腺干细胞体外诱导分化成胰岛及鉴定的技术和方法。
方法：体外获得纯化的小鼠胰腺干细胞，采用联合诱导剂对其进行成胰岛方向的诱导分化，并对诱导形成的胰岛样细胞团进行形态学观察、双硫腙染色、RT-PCR和Western blot检测。
结果与结论：实验通过细胞形态学和细胞生长特性的观察以及免疫细胞化学染色证实体外成功获得了小鼠胰腺干细胞，采用联合诱导剂将其诱导成胰岛样结构，呈球形，以较细长的蒂部与瓶底连接，双硫腙染色将其染成铁红色。RT-PCR和Western blot 法可分别检测到胰岛样细胞团的胰岛素mRNA和胰岛素蛋白。结果证实小鼠胰腺干细胞可体外诱导分化成含β细胞的胰岛样细胞团。
关键词：胰腺干细胞；诱导；胰岛样细胞团；干细胞培养；小鼠
doi:10.3969/j.issn.1673-8225.2012.10.020
BACKGROUND: Because of lacking of islet sources, the islet cells transplantation for the treatment of diabetes can not meet the clinical demand, so the differentiation of pancreas stem cells into islets in vitro has become a focus of the research.
OBJECTIVE: To in vitro induce the mice pancreas stem cells into the islet-like cell clusters and to perform the relate measurement; to investigate the techniques and methods to differentiate the pancreas stem cells into the islets as well as the detection method.  
METHODS: The mice pancreas stem cells were obtained in vitro, and then the joint inducer was used to induce the pancreas stem cells to differentiate into islets. The islet-like cell clusters were preformed with morphological observation, dithizone dyeing, RT-PCR and Western blot detection.
RESULTS AND CONCLUSION: Through the observation of cell morphology and cell growth characteristics and immunocytochemistry staining, we obtained mice pancreas stem cells in vitro. The joint inducer was used to induce the mice pancreas stem cells to differentiate into islet-like cells, the cells were spherical with a more slender pedicle connected with the bottom, and the cells was stained iron red by dithizone dyeing. The results of RT-PCR and Western blot determined the expression of insulin mRNA and insulin protein of islet-like cells respectively. It confirms that the mice pancreas stem cells can be induced in vitro to differentiate into beta containing islet-like cell clusters.