Wang Kai, Wang Xi, Han Cheng-Sheng, Chen Liang-Yi, Luo Ying
Department of Biomedical Engineering, Peking University.
Institute of Molecular Medicine, Peking University.
J Vis Exp. 2017 Jul 23(125):54995. doi: 10.3791/54995.
Islet transplantation has been clinically proven to be effective at treating type 1 diabetes. However, the current intrahepatic transplantation strategy may incur acute whole blood reactions and result in poor islet engraftment. Here, we report a robust protocol for the transplantation of islets at the extrahepatic transplantation site-the epididymal fat pad (EFP)-in a diabetic mouse model. A protocol to isolate and purify islets at high yields from C57BL/6J mice is described, as well as a transplantation method performed by seeding islets onto a decellularized scaffold (DCS) and implanting them at the EFP site in syngeneic C57BL/6J mice rendered diabetic by streptozotocin. The DCS graft containing 500 islets reversed the hyperglycemic condition within 10 days, while the free islets without DCS required at least 30 days. The normoglycemia was maintained for up to 3 months until the graft was explanted. In conclusion, DCS enhanced the engraftment of islets into the extrahepatic site of the EFP, which could easily be retrieved and might provide a reproducible and useful platform for investigating the scaffold materials, as well as other transplantation parameters required for a successful islet engraftment.
胰岛移植已在临床上被证明对治疗1型糖尿病有效。然而,当前的肝内移植策略可能会引发急性全血反应,并导致胰岛植入效果不佳。在此,我们报告了一种在糖尿病小鼠模型中,于肝外移植部位——附睾脂肪垫(EFP)进行胰岛移植的可靠方案。本文描述了一种从C57BL/6J小鼠中高产率分离和纯化胰岛的方案,以及一种通过将胰岛接种到脱细胞支架(DCS)上,并将其植入经链脲佐菌素诱导成糖尿病的同基因C57BL/6J小鼠的EFP部位的移植方法。含有500个胰岛的DCS移植物在10天内逆转了高血糖状况,而没有DCS的游离胰岛至少需要30天。正常血糖水平维持了长达3个月,直到移植物被取出。总之,DCS增强了胰岛在EFP肝外部位的植入,该部位易于获取,可能为研究支架材料以及成功进行胰岛植入所需的其他移植参数提供一个可重复且有用的平台。