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当前位置: 首页 > 文章 > 应用Gelatin和3D培养验证细胞粘附对传染性支气管炎病毒在体外培养细胞中增殖的影响 中国预防兽医学报 2018 (12) 1089-1094
Position: Home > Articles > Identification of cell adhesion as determinant of the infectious bronchitis virus replication in cell cultures using gelatin and 3D culture Chinese Journal of Preventive Veterinary Medicine 2018 (12) 1089-1094

应用Gelatin和3D培养验证细胞粘附对传染性支气管炎病毒在体外培养细胞中增殖的影响

作  者：
王娜娜;韩宗玺;山丹;邵昱昊;刘胜旺;李海
单  位：
中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽呼吸道传染病创新团队
关键词：
传染性支气管炎病毒;全基因组转录谱分析;病毒增殖;3D细胞培养
摘  要：
除传染性支气管炎病毒(IBV) Beaudette株外,其它IBV均难以在体外细胞系中传代培养。为研究限制IBV体外传代培养的影响因素,本研究应用流式细胞术检测IBV经典株M41囊膜与宿主细胞膜融合程度,结果显示M41可以在体外培养条件下侵入鸡细胞系LMH和DF-1,但荧光定量PCR检测显示侵入的M41无法增殖。进一步通过对比分析M41易感组织气管、非易感组织脾脏及LMH细胞全基因组转录谱数据,结果显示宿主细胞粘附、细胞周期、p53及SRC可能是M41在体外培养细胞中增殖的宿主决定因素。本研究通过在2D培养体系中预先孵育Gelatin和3D培养两种方式来调节细胞粘附的物理环境,结果显示两种方法均可以促进M41在体外培养细胞中的增殖;利用多种经典的细胞周期抑制剂及p53和SRC活性调控分子预先处理LMH和DF1,显示细胞周期、p53活性及SRC活性对IBV体外增殖无显著影响。表明细胞粘附特性是影响IBV体外增殖的重要因素。本研究为IBV体外细胞培养体系的建立奠定了实验基础。
译  名：
Identification of cell adhesion as determinant of the infectious bronchitis virus replication in cell cultures using gelatin and 3D culture
作  者：
WANG Na-na;HAN Zong-xi;SHAN Dan;SHAO Yu-hao;LIU Sheng-wang;LI Hai;Division of Avian Infectious Diseases, State Key Lab oratory of Veterinary Biotechnology, Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences;
单  位：
WANG Na-na%HAN Zong-xi%SHAN Dan%SHAO Yu-hao%LIU Sheng-wang%LI Hai%Division of Avian Infectious Diseases, State Key Lab oratory of Veterinary Biotechnology, Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences
关键词：
infectious bronchitis virus;;genome-wide transcriptional profile analysis;;viral replication;;3D cell culture
摘  要：
Currently,except Beaudette strain,no infectious bronchitis virus(IBV)stain can be cultured and passaged in vitro,which limits the study of the mechanism of IBV infection and pathogenesis and the development of new type of vaccine.By detecting the membrane fusion between virus and host cells via FACS,present study observed the entry of IBV stain M41 in chicken cell lines of LMH and DF1 in vitro.While subsequent viral replication was not detected in either cell lines assayed by RT-q PCR.To explore the host determinates of M41 replication,in vitro comparison of the published genome-wide transcriptional data of trachea,which is susceptible to M41 infection,and spleen and LMH cells,which are unsusceptible to M41 infection,was conducted and suggests cell adhesion,cell cycle,p53 and SRC as potential determinates of M41 replication in vitro.Modulation of cell adhesion environment by either pre-incubation of gelatin in 2D culture system or the performance of 3D cell culture promoted M41 replication.While none of cell cycle,p53 or SRC had any influence on M41 replication,as assayed by RT-qPCR in host cells pre-treated with widely-used chemical regulators of cell cycle, p53 activity and SRC activity. Our study provides fundamental data for the development of IBV culture system in vitro.

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