Reversal of diabetes in Beagle dogs by intrahepatic autotransplantation of bone-derived mesenchymal stem cells

Abstract

Objective To assess the long-time curative effect and safety of intrahepatic autotransplantation of bone-derived Beagle canine mesenchymal stem cells (BcMSCs) containing the human Insulin (INS) and enhanced green fluorescent protein (EGFP) gene through artery intervention in diabetic Beagle dogs. Methods After the double expression vector pMSCV-INS-IRES-EGFP was constructed, BcMSCs were isolated from the bone marrow of Beagle canine, expanded, and transfected with viral suspension.Then the cells were treated with G418 and cultured in vitro, followed by intrahepatic autotransplantation to the diabetic Beagle canine. Results The body weight of diabetic Beagle dogs that received BcMSCs harboring the human INS and EGFP gene stopped dropping and increased by 8.83% within 10 weeks after treatment, and the average blood glucose levels were ( 19. 80 ±3. 41 ) mmol/L (at day 7) and ( 10. 53 ±2. 12)mmol/L ( at day 70) respectively. There was significant difference in the average blood glucose level between BcMSCs group and control group (P ＜ 0. 05). The human INS was detected in blood serum and the liver of the treated animals. Intravenous glucose tolerance test revealed that in BcMSCs group the blood glucose level was reduced. Moreover, pathomorphological research confirmed that the transplanted BcMSCs were permanently planted in the animal liver and could secrete human INS. No pathomorphological changes were found in the transplant part. Conclusion Intrahepatic autotransplantation of BcMSCs expressing human INS could effectively reduce the blood glucose level and alleviate the symptoms of diabetes. Key words: Bone-derived mesenchymal stem cells; Insulin; Diabetes; Autotransplantation; Gene therapy