Muniappan Latha, Ozcan Sabire
Molecular & Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY, USA.
Methods Mol Biol. 2009;590:131-42. doi: 10.1007/978-1-60327-378-7_8.
The beta cells within the pancreatic islets are responsible for production of insulin, a peptide hormone required for maintaining normoglycemia. The establishment of efficient gene transfer into pancreatic islets is very important for studies of insulin and glucagon production and secretion, as well as for gene therapy purposes for the treatment of diabetes. We describe here in detail a protocol for adenoviral gene transfer into isolated mouse islets of the pancreas. Effective gene transfer into pancreatic islets using recombinant adenoviruses can be achieved with a multiplicity of infection (MOI) of 10. However, if the islets are not dispersed, adenoviral gene transfer is limited only to the cells on the periphery of the islets, which represent the glucagon-producing alpha cells in rodents. Dispersion of pancreatic islets with EGTA increases the efficiency of gene transfer into the cells within the core of the islets, which consist of insulin-producing beta cells.
胰岛内的β细胞负责产生胰岛素，胰岛素是维持正常血糖水平所需的一种肽类激素。建立高效的基因转移至胰岛的方法对于胰岛素和胰高血糖素产生及分泌的研究，以及糖尿病治疗的基因治疗目的而言非常重要。我们在此详细描述一种将腺病毒基因转移至分离的小鼠胰腺胰岛的方案。使用重组腺病毒实现向胰腺胰岛的有效基因转移，感染复数（MOI）为10时即可达成。然而，如果胰岛未分散，腺病毒基因转移仅局限于胰岛外周的细胞，这些细胞在啮齿动物中代表产生胰高血糖素的α细胞。用乙二醇双四乙酸（EGTA）分散胰腺胰岛可提高基因转移至胰岛核心细胞的效率，胰岛核心细胞由产生胰岛素的β细胞组成。
