超表达CD163的猪诱导性多能干细胞系构建  

Construction of Porcine(Sus scrofa)Induced Pluripotent Stem Cell Lines with Over-expression of CD163

作　　者：岳威 张炬庆 杨昕淳 吴晓龙 沈巧艳[1] 于帅 祝振硕 王承宝 张仕强 华进联[1] YUE Wei;ZHANG Ju-Qing;YANG Xin-Chun;WU Xiao-Long;SHEN Qiao-Yan;YU Shuai;ZHU Zheng-Shuo;WANG Cheng-Bao;ZHANG Shi-Qiang;HUA Jin-Lian(Shaanxi Stem Cell Engineering and Technology Research Center/College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物医学院/陕西省干细胞工程技术研究中心,杨凌712100

出　　处：《农业生物技术学报》2022年第10期2036-2044,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32072806);陕西省重点科技创新团队(2019TD-036);陕西省重点研发计划(2022NY-044)。

摘　　要：CD163作为清道夫受体蛋白,是猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)的最重要细胞受体。诱导性多能干细胞(induced pluripotent stem cells,iPSCs)是将外源转录因子导入体细胞后,经重编程获得的具有胚胎干细胞生物学特性的多能干细胞。本研究首先通过慢病毒载体系统将猪(Sus scrofa)源CD163基因转导入猪iPSCs(piPSCs),建立稳定表达CD163的诱导性多能干细胞系,将其命名为CD163OE-piPSC。病毒侵染实验证实,该细胞株对PRRSV高度易感。碱性磷酸酶染色和细胞群体倍增时间检测及5-乙炔基-2’-脱氧尿嘧啶核苷(5-ethynyl-2’-deoxyuridine,EdU)细胞增殖检测证实,CD163基因过表达不影响iPSCs的多能性与增殖能力。本研究获得的细胞株可有效应用于PPRSV与宿主互作的致病机理研究及病毒性疾病的靶标分子筛选。CD163 is a macrophage-specific protein in the cysteine superfamily rich in scavenger receptors and is the most important cellular receptor of Porcine reproductive and respiratory syndrome virus(PRRSV).Induced pluripotent stem cells(iPSCs)are pluripotent stem cells with biological characteristics of embryonic stem cells obtained by reprogramming after introducing foreign transcription factors into somatic cells.Firstly,porcine(Sus scrofa)CD163 gene was transferred into porcine induced pluripotent stem cells(piPSCs)by lentiviral vector system,and an induced pluripotent stem cell line stably expressing CD163 was established,which was named CD163OE-piPSC.The susceptibility of the cell line to PRRSV was verified by virus infection,and the pluripotency and proliferation of iPSCs were not affected by alkaline phosphatase staining,cell population doubling time test and 5-ethynyl-2’-deoxyuridine(EdU)cell proliferation test.The cell lines obtained in this study could be effectively used to study the pathogenesis of PRRSV-host interaction and screen the target molecules of viral diseases.

关 键 词：基因编辑 猪繁殖与呼吸综合征病毒(PRRSV) 诱导性多能干细胞(iPSCs) CD163 

分 类 号：S852.21[农业科学—基础兽医学]