Abstract:

Objective To investigate the labeling of mouse pancreatic islets with lentivirus Luciference and to study their fluorescence expression in vitro and in vivo after transplantation in nude mice. Methods The purified and cultured ICR mouse islets were transfected with lentivirus-LUC islet culture medium with a multiplicity of infection of 10 MOI， the islets were then washed and cultured for 24 h. The fluorescence expression of islets labeled with different isletequivalents of LUC in vitro were observed by optical imaging system of small animals within 14 days of culture. In vivofluorescence expression was observed within 30 days after renal subcapsular transplantation in nude mice. Results The activity of islets after LUC gene transfection by this method was （86.7±6.7）%. Using IVIS Lumina LT， LUC-labeled isletsshowed fluorescent expression while cultured in vitro，and the amount of fluorescent expression was related to islet equivalent. In vitro，the fluorescence expression intensity showed a trend of increasing over time within 14 days of culture. After transplanted under the renal capsule of nude mice with LUC-labeled islets， long-term fluorescence expression was seen （＞ 30 days）. However， the intensity of fluorescence expression fluctuated greatly over time.Conclusion In this study，the survival of LUC-labeled islets after transplantation in vivo can be observed in vivo for a long time， which provides a method reference for the follow-up study on the survival and efficacy of islets at the transplant site. 

Key words:

 , Luciference； , Pancreatic islets； , Nude mice； , Transplantation

摘要：

目的 探讨慢病毒 Luciference 标记小鼠胰岛并研究其在体外和裸鼠体内移植后的荧光表达。方法 以感染复数 10 MOI 的慢病毒 -LUC 胰岛培养液转染培养后的 ICR 小鼠胰岛，洗涤后再培养 24 h 后，使用小动物活体光学成像系统观察不同胰岛当量 LUC 标记后的胰岛在体外的荧光表达量，及在体外培养 14 d 内的荧光表达。在裸鼠肾被膜下移植后，观察 30 d 内的体内荧光表达情况。结果 经本方法转染 LUC 基因后的胰岛细胞团活性为（86.7±6.7）% ；在小动物活体成像仪中，体外培养的 LUC 标记胰岛可见有荧光表达，且表达量与胰岛当量有关 ；体外培养 14 d 内的荧光表达强度呈现与时间同步增强的趋势 ；LUC 标记胰岛经裸鼠肾被膜下移植后可见荧光长期表达（＞ 30 d），但表达的强度随时间变化有较大差异。结论 本研究 LUC 标记的胰岛可实现在活体状态下长期观察移植至体内胰岛的存活情况，可为后续研究胰岛在移植部位的存活和功效提供一定参考。

关键词:

荧光素酶 , 胰岛 , 裸鼠 , 移植