Xue T, Wang Y Z, Pan Q H, Wang Q, Yuan J F, Chen T S
College of Fisheries, Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, China.
Collaborative Innovation Center for Efficient and Health Production of Fisheries in Hunan Province, Changde, China.
J Fish Dis. 2018 Feb;41(2):365-374. doi: 10.1111/jfd.12736. Epub 2017 Oct 25.
In this study, we established and characterized a cell line derived from the kidney of black carp (Mylopharyngodon piceus), which is an important freshwater aquaculture species. The cell line was designated as MPK and subcultured for more than 70 passages in DMEM medium containing 10% fetal bovine serum (FBS) at 28°C. MPK had a modal diploid chromosome number of 48. Moreover, a transient MPK transfection efficiency was up to 18% using a green fluorescent protein plasmid by a modified electroporation. In addition, the MPK cells showed susceptibility to spring viremia of carp virus (SVCV), as demonstrated by the presence of severe cytopathic effects (CPEs) and increased viral RNA. Unexpectedly, the MPK cells expressed pluripotency-associated genes such as nanog, oct4 and vasa, indicating that these are possibly adult stem cells. Taken together, we have established a stable cell line from kidney that may potentially be utilized as an in vitro platform for genetic modifications and host-pathogen analysis in black carp.
在本研究中,我们建立并鉴定了一种源自草鱼(Mylopharyngodon piceus)肾脏的细胞系,草鱼是一种重要的淡水养殖物种。该细胞系被命名为MPK,并在含有10%胎牛血清(FBS)的DMEM培养基中于28°C传代培养超过70代。MPK的核型二倍体染色体数为48。此外,通过改良电穿孔法使用绿色荧光蛋白质粒,MPK的瞬时转染效率高达18%。另外,MPK细胞对鲤春病毒血症病毒(SVCV)敏感,表现为出现严重的细胞病变效应(CPEs)且病毒RNA增加。出乎意料的是,MPK细胞表达了多能性相关基因,如nanog、oct4和vasa,这表明它们可能是成体干细胞。综上所述,我们从肾脏建立了一个稳定的细胞系,其有可能被用作草鱼基因改造和宿主-病原体分析的体外平台。