Exploration of GFP labeled pancreatic islets and their transplantation in different sites of mice and rats
Objective Based on green fluorescent protein(GFP)labeling of pancreatic islets,to establish a simple and accurate method,to explore the survival effect of pancreatic islet transplantation in different parts of mice and rats.Methods GFP adenovirus plasmid transfection was used to label and isolate islets from mice and rats,and transplanted islets into mice abdominal cavity,inguinal subcutaneous tissue,and rats omentum of the same strain.After 10 d of transplantation,the distribution and survival of islets at the transplantation sites were observed with a portable excitation light source.Dithizone(DTZ)staining and fluorescence microscopy were used to confirm the pancreatic islet cells,and HE staining was used to confirm the morphology of pancreatic islet cell clusters at the transplantation sites.Results After transfection with GFP adenovirus plasmid with a multiplicity of infection of 100,the pancreatic islets were cultured for 48 h,and green fluorescence was observed under fluorescence microscopy and portable excitation light source irradiation.After transplantation of 200 GFP plasmid transfected pancreatic islets into the abdominal cavity and subcutaneous inguinal region of mice,and mesentery of rats,for 10 d,green fluorescent pancreatic islet tissue was found at the sites.Taking the subcutaneous transplantation of pancreatic islet tissue in the groin of mice as an example,after frozen sectioning with 10 μm thick and DTZ staining,pancreatic islet tissue with red color and green fluorescence expression could be seen in the tissue.By HE staining,the appearance of pancreatic islets was round.Conclusion This study established a simple and accurate method to explore the survival of islet transplantation in mice and rats,which laid a foundation for the preliminary judgment of the applicability of the transplantation site for islet transplantation and further improvement research.
green fluorescent proteinpancreatic isletsmiceratstransplantation siteislets survival
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