The Study On The Isolation And Purification Of Islets Cells From Human Pancreata And The Effects Of Immunosupperssive Agents On Islets Cells

Objectives:1.To investigate how to improve the yield and quality of human islets in islolation and purification and evaluate the quality of islet cell .2.To investigate the effects of immunosuppressive drugs on the viability of pancreatic islets cells cultured in vitro ,and to explore their effects on the outcome of ilslet transplantation.Methods:1.Human's islet cells were isolated using two times digestion by collagenase injected through pancreastic duct. Islet were purified by Ficoll-400 discontinous density gradient centrifugation and cultured. 2.The identification of islet cells and acinar cells were studied with dithizone (DTZ) and acridine orange (AO)-propidium iodide(PI). 3.Human islet cells were exposed to various concentrations of four immunosuppressive agents for 72 hr respectively. Glucose -stimulated insulin secretion during subsequent static incubation was measured using the ultrasensitive human insulin ELISA kit.Results:1.The mean yield of islet cells was 80 000 islets/pancreas.Islet purity was over 80%,islet viability was more than 95%.The insulin releasee test indicated the islet cells were sensitive to...