Wei Fulei, Hua Qiang, Liang Jian, Yue Miao, Xu Dingfan, Tian Wengen, Yu Luxian, Feng Zhaohui
State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, 251 Ningda Road, Xining, 810016, People's Republic of China.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People's Republic of China.
In Vitro Cell Dev Biol Anim. 2022 Dec;58(10):970-978. doi: 10.1007/s11626-022-00729-z. Epub 2022 Oct 26.
Gymnocypris przewalskii (naked carp), a native teleost, plays an important role in the ecosystem of Qinghai Lake (altitude, 3.2 km) on the Qinghai-Tibet Plateau in China. We developed a new cell line from the muscle of G. przewalskii using the explant technique and named the cell line GPM. This cell line was maintained in DMEM medium (high glucose) supplemented with 15% fetal bovine serum (FBS). The cell line was successfully subcultured up to 32 passages and was authenticated by immunofluorescence assay, sequencing the mitochondrial cytochrome C oxidase subunit I (COI) and 16S rRNA genes, and by chromosome analysis. In the medium containing 15% FBS, the cell line could be passaged stably at 25 °C. The GPM cell line could express green fluorescent protein (GFP) with a CMV promoter with about 5% transfection efficiency. MTT tests showed that Clostridium botulinum toxin (BTX) was toxic to the cell line. The cell line could be successfully cryopreserved in liquid nitrogen with a revival efficiency of over 70%. This study demonstrated that the GPM cell line can be used as an important tool for understanding the physiological characteristics of G. przewalskii, and it can provide a resource for studying gene function and toxicological reactions in vitro.
青海湖裸鲤是一种本地硬骨鱼，在中国青藏高原海拔3.2千米的青海湖生态系统中发挥着重要作用。我们采用组织块培养技术从青海湖裸鲤的肌肉中建立了一个新的细胞系，并将其命名为GPM。该细胞系在添加15%胎牛血清（FBS）的DMEM高糖培养基中培养。该细胞系已成功传代至32代，并通过免疫荧光分析、线粒体细胞色素C氧化酶亚基I（COI）和16S rRNA基因测序以及染色体分析进行了鉴定。在含有15%FBS的培养基中，该细胞系在25℃下可稳定传代。GPM细胞系可通过CMV启动子表达绿色荧光蛋白（GFP），转染效率约为5%。MTT试验表明，肉毒杆菌毒素（BTX）对该细胞系有毒性。该细胞系可成功保存在液氮中，复苏效率超过70%。本研究表明，GPM细胞系可作为了解青海湖裸鲤生理特性的重要工具，并可为体外研究基因功能和毒理学反应提供资源。
