Establishment and characterization of a novel cell line from midgut tissue of Helicoverpa armigera (Lepidoptera: Noctuidae)
Establishment and characterization of a novel cell line from midgut tissue of Helicoverpa armigera (Lepidoptera: Noctuidae)
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DOI：
10.1007/s11626-015-9870-6
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发表时间：
2015-02
期刊：
影响因子：
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通讯作者：
Jianghua Li;Fei He;Yongbo Yang;Yutao Xiao;Rong Peng;H. Yao;Xianchun Li;Jianxin Peng;H. Hong;Kaiyu Liu
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文献类型：
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作者：
Jianghua Li;Fei He;Yongbo Yang;Yutao Xiao;Rong Peng;H. Yao;Xianchun Li;Jianxin Peng;H. Hong;Kaiyu Liu
鳞翅目幼虫的中肠是许多病原体（例如苏云金芽孢杆菌（Bt））的目标。源自中肠组织的细胞系将成为许多研究领域非常有用的工具。然而，迄今为止，还没有 Bt 敏感的中肠来源的细胞系可用。在这里，我们报道了一种新的细胞系，命名为HNU-Ha-MG1，是从棉铃虫第四龄幼虫的中肠组织中建立的。该细胞系在补充有 10−15% 胎牛血清的 Grace 昆虫细胞培养基中生长良好。大多数细胞的形状为圆形或多边形，有的倾向于聚集形成多个细胞团。细胞直径为13.8±1.8μm，最大密度达到(2.40±0.15)×106个/ml。 28°C 时，对数生长期的群体倍增时间为 58.6±7.0 小时。染色体数目约为90-130条，表现出鳞翅目细胞系的典型染色体特征。该细胞系的随机扩增多态性DNA模式与我们实验室常用的Sl-HP和Hi5细胞系不同。 20-Hydroxyecdysterone 在 2 μg/ml 浓度下诱导极小部分细胞凋亡，但不影响自噬相关蛋白 8 (Atg8) 的表达及其在处理后 36 小时的脂化。该细胞系允许苜蓿银纹夜蛾核多角体病毒（AcMNPV）和H.棉铃虫核多角体病毒（HaSNPV）。发现该细胞系在终浓度为 0.5−1.0 μg/ml 时对活化的 Cry1C 敏感，但对活化的 Cry1Ac 不敏感。
The midgut of lepidopteran larvae serves as a target for many pathogens such asBacillus thuringiensis(Bt). Cell lines originating from midgut tissues will be very helpful tools in many research fields. However, to date, no Bt-susceptible midgut-derived cell lines are available. Here, we reported that a novel cell line, designated as HNU-Ha-MG1, was established from midgut tissue of the fourth instar larvae ofHelicoverpa armigera. This cell line grew well in Grace’s insect cell culture medium supplemented with 10−15% fetal bovine serum. The shape of the most cells was round or polygonal, and some tended to aggregate to form multiple cell masses. The size of the cells was 13.8 ± 1.8 μm in diameter, and the maximum density reached (2.40 ± 0.15) × 106cells/ml. The population doubling time during logarithmic growth phase was 58.6 ± 7.0 h at 28°C. The number of chromosomes was about 90–130, which exhibited typical chromosome characteristics of lepidopteran cell lines. The patterns of random amplified polymorphic DNA of the cell line were different from those of Sl-HP and Hi5 cell lines which were frequently used in our laboratory. 20-Hydroxyecdysterone induced apoptosis in a very small part of cells at 2 μg/ml but did not affect expression of autophagy-related protein 8 (Atg8) and its lipidation at 36 h post-treatment. The cell line was permissive toAutographa californicanuclear polyhedrosis virus (AcMNPV) andH. armigeranuclear polyhedrosis virus (HaSNPV). This cell line was found to be susceptible to activated Cry1C at the final concentration of 0.5−1.0 μg/ml but not to the activated Cry1Ac.