A new insect cell line from pupal ovary of Spodoptera exigua established by stimulation with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG)
A new insect cell line from pupal ovary of Spodoptera exigua established by stimulation with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG)
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DOI：
10.1007/s11626-012-9511-2
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发表时间：
2012-05
期刊：
影响因子：
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通讯作者：
Xuan Li;Q. Qin;Ning Zhang;Wei Zhu;Jihong Zhang;Hong-tuo Wang;L. Miao;Huan Zhang
中科院分区：
文献类型：
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作者：
Xuan Li;Q. Qin;Ning Zhang;Wei Zhu;Jihong Zhang;Hong-tuo Wang;L. Miao;Huan Zhang
用N-甲基-N'-硝基-N-亚硝基胍（MNNG）处理原代细胞，建立了甜菜夜蛾蛹卵巢的连续细胞系。用3.0μg/ml的MNNG处理细胞三天后，细胞形成单层，并在去除MNNG后最初传代培养60天，然后传代30代。建立的细胞系命名为 IOZCAS-Spex 12，由三种类型细胞的混合物组成，包括球形、纺锤形和椭圆形细胞。细胞系在对数生长期的群体倍增时间为 71 小时。 DNA扩增指纹聚合酶链式反应分析证实新细胞系来源于S.埃西瓜。研究了 IOZCAS-Spex 12 细胞对某些核多角体病毒感染的敏感性。结果表明，该细胞系对S. 感染高度敏感。外鸟核多角体病毒和苜蓿银纹夜蛾多核多角体病毒，对斜纹夜蛾核多角体病毒感染稍感，对棉铃虫核多角体病毒或美国白蛾核多角体病毒感染不易感染。这项研究的结果表明，MNNG 治疗可以克服获得持续增殖细胞的现有限制，并可能为分离的昆虫细胞永生化开辟可能性。
A continuous cell line derived from the pupal ovary ofSpodoptera exiguawas established by treating primary cells withN-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Three days after treating cells with 3.0 μg/ml of MNNG, the cells formed a monolayer and were initially subcultured 60 d after the MNNG was removed, followed by subculturing for 30 passages. The established cell line, designated IOZCAS-Spex 12, consisted of a mixture of three types of cells, including spherical, spindle-shaped, and oval cells. The population doubling time of the cell line during its logarithmic growth phase was found to be 71 h. DNA amplification fingerprinting polymerase chain reaction analysis confirmed that the new cell line originated fromS. exigua. Susceptibility of IOZCAS-Spex 12 cells to infection by certain nucleopolyhedroviruses was investigated. The results showed that the cell line was highly susceptible to infection byS. exiguanucleopolyhedrovirus andAutographa californicamultiple nucleopolyhedrovirus, slightly susceptible to infection bySpodoptera lituranucleopolyhedrovirus, and not susceptible to infection byHelicoverpa armigeranucleopolyhedroviruses orHyphantria cuneanucleopolyhedroviruses. The results of this study suggest that MNNG treatment may overcome existing limitations to obtaining continually proliferating cells and may open up the possibilities for immortalizing isolated insect cells.