Breeding of db/db mice and isolation and purification of its pancreatic islet

Objective To study the rearing method of db/db spontaneous type 2 diabetic mellitus(T2DM) mice,isolation and purification of pancreatic islet,and detection of pancreatic islet function.Method Heterozygous(+/db) were mated naturally,reared in barrier environment,and fed by complete nutritional feed.Separate the pancreas by retrograde perfusion with pancreatic duct,and purify the islets by the method of ficoll-400 density gradient centrifugation.Results The number of db/db mice accounted for 1/2 of the total breeding number,of which,the fourth birth littered the largest number of mice,and then began to decrease gradually.The purified islets under an inverted microscope were round or oval,with its purity over 90%.In vitro release test indicated the function of islet B cell was reletively poor.Conclusions db/db mice could reproduce stably in barrier environment,which could separate and purify the pancreatic islet successfully.