ISL1基因腺病毒过表达载体的构建及其诱导犬ADSCs向胰岛样细胞分化  

Construction of ISL1 adenovirus overexpression vector and its induction of canine ADSCs differentiation into islet-like cells

作　　者：张霞 戴鹏秀 高登科 阮晨梅 王璟璐 李佳锴 陈奕静 张露文 张翊华[1] ZHANG Xia;DAI Peng-xiu;GAO Deng-ke;RUAN Chen-mei;WANG Jing-lu;LI Jia-kai;CHEN Yi-jing;ZHANG Lu-wen;ZHANG Yi-hua(College of Veterinary Medicine,Northwest Agricul..ture and Forestry University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨陵712100

出　　处：《中国兽医学报》2020年第8期1515-1521,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31872529);陕西省自然科学基础研究计划重点资助项目(2019JZ-16)。

摘　　要：构建腺病毒重组表达载体pADTrack-ISL1-AdEasy,经293A细胞的包装和扩增,获得具有感染性的重组腺病毒,将其按感染复数(MOI)=100感染犬脂肪间充质干细胞(adipose-derived mesenchymal stem cells,ADSCs),观察感染后4,7,10,13,16 d绿色荧光蛋白表达情况、细胞形态变化和胰岛β细胞发育相关基因的mRNA表达情况。结果显示,含ISL1基因的重组腺病毒感染犬ADSCs 48 h后,绿色荧光蛋白开始表达;13 d后犬ADSCs出现细胞形态变化,由梭形变为神经细胞样;qRT-PCR分析表明在犬ADSCs中过表达ISL1基因提高了PDX1、NGN3、MNX1、MAFA和NKX6.1基因的内源性表达。结果表明,含ISL1基因的重组腺病毒成功感染犬ADSCs,并可促进其向胰岛样前体细胞分化,具有进一步诱导形成胰岛样细胞的可能。In order to explore the role of ISL1 gene in the differentiation of canine adipose-derived mesenchymal stem cells(ADSCs)into islet-like cells,in this study,the recombinant adenoviral expression vector pADTrack-ISL1-AdEasy was constructed,and the infectivity was obtained by packaging and amplification of 293 A cells.The canine ADSCs were infected with the infection number MOI=100.The expression of green fluorescent protein,cell morphology and mRNA expression of isletβcell development related genes were observed at 4,7,10,13,16 d.The results showed that green fluorescent protein begin to express in canine ADSCs at 48 hours after infection;after 13 d,the morphology of canine ADSCs changed from fusiform to neuron-like;qRT-PCR analysis showed that in canine ADSCs,overexpression of the ISL1 gene increased endogenous expression of PDX1,NGN3,MNX1,MAFA,NKX6.1.The results confirmed that the recombinant adenovirus containing ISL1 gene successfully infected canine ADSCs and promoted its differentiation into islet-like precursor cells,which may further induce the formation of islet-like cells.

关 键 词：ISL1基因 腺病毒载体 犬ADSCs 胰岛样细胞 

分 类 号：S852.2[农业科学—基础兽医学] S852.65[农业科学—兽医学]