Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
| 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
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| 2018-11-01 | |
| 发表期刊 | DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
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| ISSN | 0145-305X |
| 卷号 | 88页码:45-54 |
| 通讯作者 | Xiang, Jianhai([email protected]) |
| 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 |
| DOI | 10.1016/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]###2224; Qingdao National Laboratory for Marine Science and Technology[2015ASKJ02]###792; 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 | |
| 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. |
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