Khatri Rahul, Hussmann Birte, Rawat Divya, Gürol Ali Osman, Linn Thomas
Third Medical Department, Clinical Research Lab.
Istanbul University Department of Immunology, Institute of Experimental Medicine (DETAE).
J Vis Exp. 2018 May 5(135):57559. doi: 10.3791/57559.
Pancreatic islet transplantation to reduce hyperglycemia is highly successful in rodents with chemically-induced diabetes. The most common transplantation site in experimental islet transplantation is the kidney capsule. However, as less is known about the interaction of pancreatic islets with blood constituents, it also makes sense to utilize the portal vein approach in experimental islet transplantation. This protocol demonstrates an intraportal islet transplantation technique in NMRI nude mice. Streptozotocin (180 mg/kg) is injected intraperitoneally to induce hyperglycemia in recipient mice. They are considered as diabetic at a non-fasting blood glucose level greater than 20 mmol/L. One day prior to transplantation, mouse pancreatic islets are isolated from the donor pancreas by collagenase digestion; a minimum of 350 islets are utilized per diabetic recipient. Depending upon the islet isolation yield, two or more donor mice are utilized per recipient. After overnight culture at 37 °C, islets are administered into the recipient liver via the portal vein. After surgery, the mice are protected in red Makrolon houses and observed until are awake. This protocol maintains glycemic control for 120 days in syngeneic mice and 15 days in allogeneic mice.
在化学诱导糖尿病的啮齿动物中,胰腺胰岛移植以降低高血糖症非常成功。实验性胰岛移植中最常见的移植部位是肾被膜。然而,由于对胰岛与血液成分之间的相互作用了解较少,在实验性胰岛移植中采用门静脉途径也是合理的。本方案展示了一种在NMRI裸鼠中的门静脉内胰岛移植技术。腹腔注射链脲佐菌素(180mg/kg)以诱导受体小鼠出现高血糖症。当非空腹血糖水平大于20mmol/L时,它们被视为糖尿病小鼠。在移植前一天,通过胶原酶消化从供体胰腺中分离小鼠胰腺胰岛;每个糖尿病受体至少使用350个胰岛。根据胰岛分离产量,每个受体使用两只或更多供体小鼠。在37°C过夜培养后,通过门静脉将胰岛注入受体肝脏。手术后,将小鼠置于红色聚碳酸酯饲养笼中保护,观察直至苏醒。该方案在同基因小鼠中维持血糖控制120天,在异基因小鼠中维持15天。