小鼠胰腺干细胞的分离方法。

Method for isolation of mouse pancreatic stem cells.

作者信息

Noguchi H, Matsumoto S, Ueda M, Hayashi S, Kobayashi N, Jackson A, Naziruddin B, Levy M F

机构信息

Baylor Institute for Immunology Research/Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, Texas 75204, USA.

出版信息

Transplant Proc. 2008 Mar;40(2):422-3. doi: 10.1016/j.transproceed.2008.01.007.

DOI:10.1016/j.transproceed.2008.01.007

PMID:18374088

Abstract

Replacement of beta-cell mass offers an alternative to standard insulin treatment for diabetes and may overcome the long-term side effects associated with current therapies. Pancreatic stem/progenitor cells could become a useful target for beta-cell replacement therapy in diabetic patients. We have established a method for isolating mouse pancreatic stem cells. In this study, pancreatic stem cells were isolated from 8-week-old mice. After purification on a density gradient, the density range of 1.062-1.11 contained pancreatic stem cells. The islets from the layers were deleted by dithizone staining and hand-picking under a dissecting microscope. The remnant cells were then cultured, inoculated into 96-well plates, and cloned by limiting dilution. One of the wells contained cells, named HN#5 cells, which expressed ductal cell markers, such as cytokeratin-19. HN#5 cells differentiated into insulin-producing cells and albumin-producing cells by induction medium. The isolation technique described here may be useful for identification and isolation of human pancreatic stem/progenitor cells.

摘要

替换β细胞团为糖尿病的标准胰岛素治疗提供了一种替代方案，并且可能克服当前疗法相关的长期副作用。胰腺干/祖细胞可能成为糖尿病患者β细胞替代疗法的一个有用靶点。我们已经建立了一种分离小鼠胰腺干细胞的方法。在本研究中，从8周龄小鼠中分离出胰腺干细胞。在密度梯度上纯化后，密度范围为1.062 - 1.11的部分含有胰腺干细胞。通过双硫腙染色并在解剖显微镜下手工挑选，去除各层中的胰岛。然后将剩余细胞进行培养，接种到96孔板中，并通过有限稀释法进行克隆。其中一个孔中的细胞命名为HN#5细胞，其表达导管细胞标志物，如细胞角蛋白-19。HN#5细胞通过诱导培养基分化为胰岛素分泌细胞和白蛋白分泌细胞。这里描述的分离技术可能有助于鉴定和分离人胰腺干/祖细胞。