Research on primary culture model of pancreatic islets in rats / 中国临床药理学与治疗学

ABSTRACT

AIM: To develop a simple, convenience, and inexpensive method on primary culture model of pancreatic islets in rats for the study of anti-diabetic drugs. METHODS: The pancreases of SD rats were separated from the pancreatic duct with cold Hank' s solution and picked. Then the pancreases were cut into pieces and repeatedly digested by collagenase at 37℃ for the short durations of the experiment. The isolated islets were identified 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-pernate and cell lysate and the experiment of insulin secretion by stimulation of glucose and implantation of micewith 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, secretion 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. CONCLUSION: The islets got in this study have higher purity and viability with the normal 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 .