Protective effect of pancreatic stem cells in vitro on the islets of langerhans

ZHANG Yu-sen; CHANG Cai-hong; ZHANG Yue; LI Fu-rong; BAO Shi-yun

doi:10.3969/j.issn.1002-266X.2014.15.005

Abstract：Objective To investigate the protective effect of pancreatic stem cells ( PSCs) of fetal rats in vitro on the islets of langerhans.Methods The PSCs of SD rat embryos on embryonic 16th day were isolated, purified, cultivated and passaged , then were identified by immuocytochemistry and flow cytometry .The rat's islets were isolated from the pancreas . After being identified by dithizone , the islets were divided into two groups:group A which underwent islet alone culturing and group B which underwent co-culturing of islet and PSCs for 14 days.The morphological changes , survival rate and ap-optosis rate of islets were observed under inverted microscope , fluorescence microscope and flow cytometry ( FCM) in both groups.The amount of insulin secretion was measured by ELISA;and insulin stimulation index was calculated with the in-sulin release test.After being cultured for 7 days, the islets in the two groups were transplanted to the peplos of the left kid-ney in SD rats with diabetes , respectively .The blood glucose of the SD rats was tested after transplantation .Results The immuocytochemistry showed the PSCs were nestin-positive after being cultivated and passaged for the 3rd generation.Flow cytometry showed its content accounted for 74.1%;on the 7th, 14th day, the survival rate of islets in group B was signifi-cantly higher than that in group A (all P<0.01), while on the 7th day, the apoptosis rate of islets in group B was signifi-cantly lower than that in group A (P<0.05).On the 7th , 14th day, under the high glucose stimulation , the levels of in-sulin secretion and insulin stimulation index in group B were significantly higher than those in group A (all P<0.01).Af-ter transplantation in group B , the blood glucose of rats dropped to normal on the 5th day.Conclusion Co-cultivation of islet and PSCs can significantly prolong the survival time of islets and preserve viability of islets .