Isolation And Cloning Of Fetal Porcine Pancreatic Stem Cell

Diabetes mellitus is a devastating disease suffered by about 150 million people in the world. At present, this disease could not cured completely by the therapy of traditional drugs or exogenous insulin injection. Along with the transplantation success of pancreatic islets and the improvement of immunosuppressive regimen, the possibilities of curing the type I and some of type II diabetes become more and more promising. However, the widespread application of this transplant technology has greatly been restricted by the deficient human pancreatic islets from the cadavers.In order to exert islet transplantation people turn their view to heterogeneity-porcine.The best xenogenic-animal surrogate pancreatic stem cells is the pancreatic stem cells of fetal piglets, it has many advantages which possess of unlimited donor-tissue source because high breeding, similar structure and bioactive with human being, approximative blood-glucose regulation-point with anthropobiology, surviving in human serum harmLess to body, therefore, the study on pancreatic stem cells of fetal piglets is becoming the hopeful cure of diabetes. This dissertation focuses on the study of fetal porcine pancreatic stem cells from which are the isolation and cultivation of fetal pancreatic stem cells,. the characterization of biology in pancreatic cells of fetal porcine , and the multipotents via inducing and differentiating. The basal data laid a foundation of clinical applications.1 This work based on the previous work in our laboratory,found that the most suitable medium and matrix for fetal porcine duct-derived pancreatic stem cell adhering and cultue was M199 and glatin.Purified the fetal porcine duct-derived pancreatic stem cell by the method of digesting and adhering,which was positive for CK-19 and PDX-1.In this study some of the primary culture cells were positive for the ESC marker OCT-4,but they lost in the process of subculture.The purified fetal porcine duct-derived pancreatic stem cells have good proliferate ability in vitro,when were induced by Nic and HGF these cells can differentiated into pancreatic endocrine cells and excrete insulin.The insulin secretion quantity reach the maximum in the 12-15d which proved that the cell reach the most mature stage,cells will die more if induced for more time.Fetal porcine duct-derived pancreatic stem cell cannot be induced to differentiate into smooth muscle,neuron or osteoblasts,which showed that these cell did not have the pluripotency or the method was not suitable for these cells.2 Adopt the method of Zulewski et al isolated the islet-derived fetal porcine pancreatic...