CAJ | 학술논문

目的改进小鼠胰岛分离技术,结合肾被膜下移植,探讨一套完整的小鼠胰岛移植模型方案.方法结扎小鼠胆总管末段,2 g/L胶原酶V3 ml灌注胰腺,100μm网筛过滤,显微镜下使用手检法获得胰岛,对胰岛的产量、纯度、体外分泌胰岛素的功能和移植效果进行评估,使用传统的Ficoll梯度离心纯化胰岛法所获得的胰岛作为对照.结果使用改良法平均每只小鼠可分离出110个胰岛,明显多于对照组(P＜0.01),但胰岛的纯度无明显差异,改良法所分离的胰岛体外分泌胰岛素的能力强于对照组(P＜0.01),且逆转高血糖移植胰岛的量少于对照组.结论使用小鼠胰岛分离改良法能够获得更大产量和胰岛素分泌功能更好的胰岛,其纯度没有受到影响,改良法较Ficoll法省时、省材,结合肾被膜下移植所构建的胰岛移植模型可以广泛用于小鼠胰岛移植的研究.
목적 개진소서이도분리기술,결합신피막하이식,탐토일투완정적소서이도이식모형방안.방법 결찰소서담총관말단,2 g/L효원매V3 ml관주이선,100μm망사과려,현미경하사용수검법획득이도,대이도적산량、순도、체외분비이도소적공능화이식효과진행평고,사용전통적Ficoll제도리심순화이도법소획득적이도작위대조.결과 사용개량법평균매지소서가분리출110개이도,명현다우대조조(P＜0.01),단이도적순도무명현차이,개량법소분리적이도체외분비이도소적능력강우대조조(P＜0.01),차역전고혈당이식이도적량소우대조조.결론 사용소서이도분리개량법능구획득경대산량화이도소분비공능경호적이도,기순도몰유수도영향,개량법교Ficoll법성시、성재,결합신피막하이식소구건적이도이식모형가이엄범용우소서이도이식적연구.
Objective To present a series of islet transplantation in mice by improve the method of islet isolation and subcapsular kidney transplantation.Methods Islet isolation was completed by end of the common bile duct ligation,Perfusion of the pancreas by 3 ml collagenase V (2 g/L),filtration by 100 μm filter and purification by picking islets under microscopy.Islets' yield,purity,function of insulin secretion and effect of islet transplantation were evaluated compared with islets isolated by Ficoll density gradient separation method.Results Purification by filtration increased islet yield (P ＜ 0.01),and resulted in higher insulin secretion in vitro (P ＜ 0.01).Moreover,when syngeneic diabetic hosts were transplanted with a marginal islet mass,the number of islets purified by filtration which restore normoglycemia significantly was less than those isolated by Ficoll.Conclusion Islet filtration is a simple and rapid procedure for purification of islets that demonstrate improved functional mass.The islet transplantation model consisted by Islet filtration and subcapsular kidney transplantation might be widely used in the research of islet transplantation.