Survival and proliferation of pancreatic islets isolated from adult mouse in laboratory culture conditions

Recent advances in directed differentiation of pancreatic stem cells offers potential to the development of replacement therapy for diabetic patients. However, the existing differentiation protocols are complex, time-consuming, and costly; thus there is a need for alternative protocols. Today, co-culturing with pancreatic islets is apparently a promising protocol for producing beta cells. Furthermore, we need keep islets viable in vitro for extended time periods.
We maintained isolated pancreatic islets obtained from the mouse pancreas in tissue culture for 2 weeks, after which we studied the viability, proliferation and morphology of the islets. Pancreatic islets were isolated from overnight-fasted male NMRI mice by Lacy and Kostianovsky modified collagenase digestion method and islets were tested for their specificity by dithizone (DTZ) staining. Also, islet cell viability was tested by MTT assay.
Our results showed that viable pancreatic islets can be isolated from the pancreas of adult mice and maintained in tissue culture for at least 1 week, without loss of the specific functions of the cells. Cell viability of pancreatic islets was decreased after one week and also, the cell number decreased over time. Cell viability and cell number were decrease if cells were incubated for long time.
It remains to be established whether such islets will survive and remain functionally competent after co culturing. Survey of viability of pancreatic islets is necessary in in vitro, because these were used for cell therapy for diabetes.