不同体重供体及理化因素对小鼠胰岛分离的影响

Effects of the weight of the donors and physical and chemical factors on mouse pancreatic islet isolation

目的:探讨提高小鼠胰岛数量和质量的可靠方法。方法:采用胆管内胶原酶灌注和Histopaque1077密度离心法分离纯化胰岛。分别观察供体的体重、胶原酶的浓度、消化的时间及温度对胰岛分离起的作用。在体外,ELISA检测葡萄糖刺激引起的小鼠胰岛素分泌。在体内,胰岛移植评价胰岛对糖尿病小鼠血糖的调节功能。结果:高体重供体组胰岛的收获量显著高于低体重供体组(P<0.01)。胶原酶浓度、消化时间对胰岛收获量起重要作用。胰岛活率≥95%。体外胰岛素释放试验表明胰岛功能良好。同种异体胰岛移植的糖尿病小鼠高血糖状态得到逆转。结论:小鼠的体重、胶原酶的浓度、消化时间和温度是影响胰岛数量和功能的重要因素。

Objective： To study the methods and influencing factors for efficacious isolation and transplantation of mouse pancreatic islets. Methods： Islets were digested by pancreatic ductal injection of collagenase and purified by histopaque 1 077 density centrifugation method. Glucose-stimulated insulin secretion test and islet transplantation were performed to evaluate the function of isolated islets in vitro and in vivo. Results： Mice with a higher weight showed a higher islet yield than those with a lower weight （P 〈 0.01 ）. Islet viabilities were more than 95%. Islet yield varied with the application of different concentration of collagenase, the time and temperature during digestion. The function of isolated islets was good in insulin secretion both in vitro and in vivo. Conclusion： The results indicate that the weight of the mouse donors, the concentration of collagenase, and the time and temperature of digestion are important factors for efficacious isolation of islets.