Study On The Coculture Of Sertoli Cells And Islets In Mouse

In recent years, the diabetes mellitus falls ill a rate to hoick all over the world, becoming the main disease to threat mankind's health and life safety.How overrun this hard nut to crack in the world, the cancellation sufferer's pain has already become a global problem gradually.Studied on it all the way at home and abroad, in addition to medicine therapy,the islet transplantation is one of the valid methods to cure insulin dependence diabete,so become the point of domestic and international research.How isolate and purify enough the best quality islet, extension islet life time in vitro and removing immunoreaction after islet transplant, is one big topic of treating the diabetes by islet transplant currently. Sertoli cell is the key cell that support sperm development in the testicle tissue, has nourishment in the meantime, support, secrete, gulp, exempts from immunity etc.Aim at currently the Islet transplantation and sertoli cell present research condition,this research mastered and optimized the methods of isolation,purification and cultured of mouse islet and sertoli cell, then cocultured the different amount sertoli cell and islet under appropriate condition,in order to investigate sertoli cell provide new tiny environment for islet, keeping the good function of islet, extension islet life time in vitro, thus to estabish theories foundation for islet transplant applying widely.Adopted machine isolation and collagenase two steps digest method to carry on isolation to mouse islet, and then filtrated with filter net ,and Ficoll-400 density grads centrifugation, transformed petri dish after 24 hs to purify cultured islet,observed islet configuration with the fluorescence microscope, measured the insulin secrete quantity with fluorescence immunity method .Adopted trypsin andcollagenase steps digest method to isolate sertoli cell from the mouse testicle,then carried on a low infiltrattive treatment to isolated cells with the 1/3 Hank's low infiltrattive liquid, after cultured 24 hs took a purification treat with 50mmol/L Tris-HCL (PH 7.1 ), observed sertoli cell configuration and biology characteristic with HE colouration and oil red O colouration method.Cocultured the different amount sertoli cell and islet,observed islet configuration with the fluorescence microscope,measured the insulin secrete quantity and amylopsin content with fluorescence immunity and colorimetry methods.The result indicated:1. After isolation and purification, each average mouse could acquire 642±57 islet (IEQ unit), the diameter of 85% around islet was between the 100～300μm, which could...