绵羊成纤维细胞OAR-L1高效转染方法的探索

S855.9; 为了在绵羊肺成纤维细胞OAR-L1中高效表达外源蛋白,探索适合该细胞的转染方法,本研究比较了聚乙烯亚胺(polyethyleneimine,PEI)、脂质体2000转染试剂(LipofectamineTM 2000 transfection reagent,Lipo 2000)、成纤维细胞转染试剂盒(CytofectTM fibroblast transfection kit,CF2)以及慢病毒介导的细胞转染方法在OAR-L1细胞中表达外源蛋白的效果.结果显示:以pLentiCMV-EGFP-Puro或pLentiCMV-mCherry-Puro荧光质粒转染OAR-L1细胞时,...

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Published in浙江大学学报（农业与生命科学版） Vol. 48; no. 1; pp. 96 - 105
Main Authors 吴飞, 吴杰, 陈学秋, 周静茹, 张惠, 黄艳, 时恒枝, 杨怡, 马光旭, 杜爱芳
Format Journal Article
LanguageChinese
Published 浙江大学动物科学学院动物预防医学研究所,杭州 310058 01.02.2022
Subjects
成纤维细胞OAR-L1;转染方法;慢病毒;绵羊
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ISSN1008-9209
DOI10.3785/j.issn.1008-9209.2021.01.192

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Abstract S855.9; 为了在绵羊肺成纤维细胞OAR-L1中高效表达外源蛋白,探索适合该细胞的转染方法,本研究比较了聚乙烯亚胺(polyethyleneimine,PEI)、脂质体2000转染试剂(LipofectamineTM 2000 transfection reagent,Lipo 2000)、成纤维细胞转染试剂盒(CytofectTM fibroblast transfection kit,CF2)以及慢病毒介导的细胞转染方法在OAR-L1细胞中表达外源蛋白的效果.结果显示:以pLentiCMV-EGFP-Puro或pLentiCMV-mCherry-Puro荧光质粒转染OAR-L1细胞时,细胞密度和转染试剂用量均可影响PEI、Lipo 2000和CF2介导的细胞转染效率,且三者最佳转染效率均不超过30％,而慢病毒介导的细胞侵染不受细胞密度和病毒液用量的限制,获得的荧光细胞数目也显著高于前三者.包装好的病毒液可以在4或-80℃条件下至少保存15 d而侵染效果不受影响.在OAR-L1细胞中共表达2种外源蛋白时,包装获得的2种慢病毒液等比例混合后再侵染的方式能获得更高的共转率.综上所述,慢病毒侵染是一种能够实现在OAR-L1细胞中高效表达外源蛋白的细胞转染方式,本研究结果为其他难转染细胞系转染方式的选择提供了一定的参考.
AbstractList S855.9; 为了在绵羊肺成纤维细胞OAR-L1中高效表达外源蛋白,探索适合该细胞的转染方法,本研究比较了聚乙烯亚胺(polyethyleneimine,PEI)、脂质体2000转染试剂(LipofectamineTM 2000 transfection reagent,Lipo 2000)、成纤维细胞转染试剂盒(CytofectTM fibroblast transfection kit,CF2)以及慢病毒介导的细胞转染方法在OAR-L1细胞中表达外源蛋白的效果.结果显示:以pLentiCMV-EGFP-Puro或pLentiCMV-mCherry-Puro荧光质粒转染OAR-L1细胞时,细胞密度和转染试剂用量均可影响PEI、Lipo 2000和CF2介导的细胞转染效率,且三者最佳转染效率均不超过30％,而慢病毒介导的细胞侵染不受细胞密度和病毒液用量的限制,获得的荧光细胞数目也显著高于前三者.包装好的病毒液可以在4或-80℃条件下至少保存15 d而侵染效果不受影响.在OAR-L1细胞中共表达2种外源蛋白时,包装获得的2种慢病毒液等比例混合后再侵染的方式能获得更高的共转率.综上所述,慢病毒侵染是一种能够实现在OAR-L1细胞中高效表达外源蛋白的细胞转染方式,本研究结果为其他难转染细胞系转染方式的选择提供了一定的参考.
Author 吴飞
陈学秋
杨怡
吴杰
周静茹
张惠
时恒枝
黄艳
杜爱芳
马光旭
AuthorAffiliation 浙江大学动物科学学院动物预防医学研究所,杭州 310058
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Author_FL DU Aifang
ZHANG Hui
CHEN Xueqiu
MA Guangxu
SHI Hengzhi
WU Jie
HUANG Yan
WU Fei
ZHOU Jingru
YANG Yi
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Snippet S855.9; 为了在绵羊肺成纤维细胞OAR-L1中高效表达外源蛋白,探索适合该细胞的转染方法,本研究比较了聚乙烯亚胺(polyethyleneimine,PEI)、脂质体2000转染试...
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Title 绵羊成纤维细胞OAR-L1高效转染方法的探索
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