关键词: 大鼠, 胰岛细胞, 原代培养, 细胞模型, 糖屎病

Abstract: AIM: To develop a simple, convenience, and inexpensive method on primary culture model of pan-creatic islets in rats for the study of anti-diabetic drugs. METHODS: The pancreases of SD rats were separated from the pancreatic duct with cold HankJ s solution and picked. Then the pancreases were cut into pieces and re-peatedly digested by collagenase at 37 ℃ for the short du-rations of the experiment. The isolated islets were identi-fied by dithizone staining and the viability was evaluated by trypan blue staining. Pancreatic islets were incubated in RPMI 1640 or DMEM for 14-16 h, then they were transferred to new culture plates with the same medium mentioned above. Determination of insulin content of su-pemate and cell lysate and the experiment of insulin se-cretion by stimulation of glucose and implantation of mice with STZ-induced diabetes were used for evaluated the function of islets. RESULTS: The viability of isolated pancreatic islets was more than 95% and the purity of cultured islets was about 85 %. The insulin synthesis, se-cretion and sensitivity of islets stimulate by glucose which were cultured in RPMI 1640 were higher than that in DMEM. The levels of blood glucose recovered to normal in type 1 diabetic mice after islets implantation. CON-CLUSION: The islets got in this study have higher purity and viability with the nonnal biological activity for about 7 days by this method and they can be used as a cell model for the study of diabetes in vitro.

Key words: rats, pancreatic islets, primary culture, cell model, diabetes