Development of a primary culture system for haematopoietic tissue cells from Cherax quadricarinatus and an exploration of transfection methods

doi:10.1016/j.dci.2018.07.006

CORC > 海洋研究所 > 中国科学院海洋研究所 > 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, Xiaohui 1,2; Duan, Hu 1,2; Shi, Yingli1,2 ; Xie, Shijun1,2 ; Song, Zhan 3; Jin, Songjun 1,2; Li, Fuhua 1,2; Xiang, Jianhai1,2
刊名	DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
	2018-11-01
卷号	88 页码:45-54
关键词	Cherax quadricarinatus Culture of haematopoietic tissue cells Transfection WSSV infection
ISSN号	0145-305X
DOI	10.1016/j.dci.2018.07.006
通讯作者	Xiang, Jianhai(jhxiang@qdio.ac.cn)
英文摘要	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.
资助项目	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
语种	英语
出版者	ELSEVIER SCI LTD
WOS记录号	WOS:000443668600006
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/160271]
专题	海洋研究所_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
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.