猪巨噬细胞培养模型的建立与应用  

Establishment and application of a porcine macrophage culture model

作　　者：冯春燕[1,2] 于浩洋 史喜菊 杨振 王彩霞[1] 王晶晶[1] 吴绍强 林祥梅[1] FENG Chunyan;YU Haoyang;SHI Xiju;YANG Zhen;WANG Caixia;WANG Jingjing;WU Shaoqiang;LIN Xiangmei*(Chinese Academy of Inspection and Quarantine,Beijing,100176,China;CAIQ Center for Biosafety in Sanya;Science and Technology Research Center of China Customs;Nanjing Agricultural University)

机构地区：[1]中国检验检疫科学研究院动物检验与检疫研究所,北京100176 [2]三亚中国检科院生物安全中心 [3]中国海关科学技术研究中心 [4]南京农业大学

出　　处：《质量安全与检验检测》2023年第3期6-11,共6页QUALITY SAFETY INSPECTION AND TESTING

基　　金：“十四五”国家重点研发计划项目(2022YFD1800500)。

摘　　要：本研究建立了猪肺泡巨噬细胞(porcine alveolar macrophage,PAM)的体外细胞培养模型,并利用该模型进行了病毒及遗传学操作等方面的初步应用。分离猪肺泡巨噬细胞后,利用多种培养条件进行培养和观察,通过光学显微镜对培养后的PAM进行形态学观察,并利用脂多糖诱导其分化,检测分泌细胞因子的能力,建立PAM细胞体外培养模型。为进一步验证PAM细胞模型的功能,利用对表达红色荧光蛋白的大肠埃希菌(E.coli)检测PAM的吞噬功能;利用Porcine Reproductive and Respiratory Syndrome Virus(PRRSV)疫苗毒鉴定其扩繁病毒的能力;利用体外包装的慢病毒对PAM细胞进行基因操作,并对慢病毒感染前后病毒的吞噬功能进行研究。结果显示,本研究建立PAM细胞培养模型,细胞生长状态良好且对E.coli有强的吞噬能力;病毒感染实验显示,诱导产生的猪PAM细胞能有效的扩繁PRRSV疫苗毒;体外包装的慢病毒也能感染本研究的细胞模型,且感染后PAM细胞仍然对细菌具备吞噬功能。本研究成功建立了PAM细胞体外培养模型,为研究PAM细胞的免疫分化功能、病毒的感染免疫细胞的免疫机制等提供了坚实的研究基础。This study aims to establish a culture model of porcine alveolar macrophage(PAM)cells.Then,the PAM cell cultural model is further applicated and utilized.After isolating porcine alveolar macrophages,three kinds of culture conditions were used.The morphology of PAM induced by differentiation was observed using an optical microscope.To further validate the function of the PAM cell model,the phagocytic function of PAM was detected.Identification the ability of PRRSVvaccine to proliferate the virus;The Lentivirus packaged in vitro was used for gene manipulation of PAM,and the phagocytic function of the virus before and after Lentivirus infection was studied.The results showed that this study established a PAM cell culture model,with good cell growth status and good phagocytic ability against Escherichia coli;Virus infection experiments have shown that the induced pig macrophages can effectively proliferate PRRSV vaccine virus;Lentivirus packaged in vitro can also infect the cell model in this study,and PAM cells still have phagocytic function to bacteria after infection.This study has successfully established the culture model of PAM cells in vitro,which would pave the way for the further study of immune cell differentiation and the interaction between pathogenic microorganisms and immune cells.

关 键 词：巨噬细胞 细胞培养模型 吞噬功能 慢病毒 

分 类 号：S852.4[农业科学—基础兽医学]