IOCAS-IR  > 实验海洋生物学重点实验室
Development of a primary culture system for haematopoietic tissue cells from Cherax quadricarinatus and an exploration of transfection methods
Xu, Xiaohui1,2; Duan, Hu1,2; Shi, Yingli1,2; Xie, Shijun1,2; Song, Zhan3; Jin, Songjun1,2; Li, Fuhua1,2; Xiang, Jianhai1,2
2018-11-01
发表期刊DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
ISSN0145-305X
卷号88页码:45-54
通讯作者Xiang, Jianhai(jhxiang@qdio.ac.cn)
摘要Various known and unknown viral diseases can threaten crustacean aquaculture. To develop prophylactic and therapeutic strategies against viruses, crustacean cell lines are urgently needed for immunology and virology studies. However, there are currently no permanent crustacean cell lines available. In this study, we developed a new method for preparing crayfish plasma (CP) and found that CP enhanced the proliferative capacity of haematopoietic tissue (hpt) cells from Cherax quadricarinatus by an EdU (5-ethynyl-2'-deoxyuridine) assay. The optimal CP concentration for hpt cell culture and the optimal subculture method are discussed. To achieve efficient expression of a foreign gene in hpt cells cultured in vitro, different transfection methods and vectors were analysed. We found that Lipofectamine 2000 could be used to efficiently transfect a foreign vector into hpt cells and exhibited a lower level of cytotoxicity than the other methods tested, and transfection of pEGFP-N1/w249 and pDHsp70-EGFP-FLAG resulted in high EGFP expression. By transmission electron microscopy (TEM) and virus copy number analysis, we found that white spot syndrome virus (WSSV) could infect hpt cells and multiply efficiently. Our results implied that the crayfish hpt cell culture system we improved could be used as a replacement for immortal crustacean cell lines in viral infection studies. Our findings provide a solid foundation for future immortalization and gene function studies in crustacean cells.
关键词Cherax quadricarinatus Culture of haematopoietic tissue cells Transfection WSSV infection
DOI10.1016/j.dci.2018.07.006
收录类别SCI
语种英语
资助项目Major State Basic Research Development Programs of China (973 Program)[2012CB114403] ; Qingdao National Laboratory for Marine Science and Technology[2015ASKJ02] ; Major State Basic Research Development Programs of China (973 Program)[2012CB114403] ; Qingdao National Laboratory for Marine Science and Technology[2015ASKJ02]
WOS研究方向Fisheries ; Immunology ; Zoology
WOS类目Fisheries ; Immunology ; Zoology
WOS记录号WOS:000443668600006
出版者ELSEVIER SCI LTD
引用统计
被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/160272
专题实验海洋生物学重点实验室
通讯作者Xiang, Jianhai
作者单位1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, 7 Nanhai Rd, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China
3.Ocean Univ China, Coll Marine Life Sci, Qingdao, Peoples R China
第一作者单位中国科学院海洋研究所
通讯作者单位中国科学院海洋研究所
推荐引用方式
GB/T 7714
Xu, Xiaohui,Duan, Hu,Shi, Yingli,et al. Development of a primary culture system for haematopoietic tissue cells from Cherax quadricarinatus and an exploration of transfection methods[J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2018,88:45-54.
APA Xu, Xiaohui.,Duan, Hu.,Shi, Yingli.,Xie, Shijun.,Song, Zhan.,...&Xiang, Jianhai.(2018).Development of a primary culture system for haematopoietic tissue cells from Cherax quadricarinatus and an exploration of transfection methods.DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,88,45-54.
MLA Xu, Xiaohui,et al."Development of a primary culture system for haematopoietic tissue cells from Cherax quadricarinatus and an exploration of transfection methods".DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 88(2018):45-54.
条目包含的文件
文件名称/大小 文献类型 版本类型 开放类型 使用许可
Development of a pri(6674KB)期刊论文出版稿限制开放CC BY-NC-SA浏览
个性服务
推荐该条目
保存到收藏夹
查看访问统计
导出为Endnote文件
谷歌学术
谷歌学术中相似的文章
[Xu, Xiaohui]的文章
[Duan, Hu]的文章
[Shi, Yingli]的文章
百度学术
百度学术中相似的文章
[Xu, Xiaohui]的文章
[Duan, Hu]的文章
[Shi, Yingli]的文章
必应学术
必应学术中相似的文章
[Xu, Xiaohui]的文章
[Duan, Hu]的文章
[Shi, Yingli]的文章
相关权益政策
暂无数据
收藏/分享
文件名: Development of a primary culture system for haematopoietic tissue cells from Cherax quadricarinatus and an exploration of transfection methods.pdf
格式: Adobe PDF
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。