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适应在37摄氏度生长的草地贪夜蛾昆虫细胞系:一个内二倍体克隆的特性分析

Adaptation of an insect cell line of Spodoptera frugiperda to grow at 37 degrees C: characterization of an endodiploid clone.

作者信息

Gerbal M, Fournier P, Barry P, Mariller M, Odier F, Devauchelle G, Duonor-Cerutti M

机构信息

Laboratoire de Pathologie Coanparée, INRA-CNRS URA 2209, Saint-Christol-lèz-Alès, France.

出版信息

In Vitro Cell Dev Biol Anim. 2000 Feb;36(2):117-24. doi: 10.1290/1071-2690(2000)036<0117:aoaicl>2.0.co;2.

Abstract

Sf21 and Sf9 cell lines established from the lepidoptera Spodoptera frugiperda do not display major induction of heat shock proteins when exposed to a temperature of 37 degrees C. After some months of adaptation at 37 degrees C we obtained two new cell lines, Sf21-HT and Sf9-HT, which have now been established for several years in our laboratory. The Sf9-HT line displays a slightly shorter doubling time at 37 degrees C than the wild type at 28 degrees C, but cell lethality gives rise to an earlier growth arrest. The composition of total lipid extract from heat-adapted cells reveals a higher sphingomyelin to phosphatidylcholine ratio and a higher percentage of saturated fatty acids, which are expected for the lower membrane fluidity, required for thermotolerance. The cell volume of Sf9-HT is doubled, and by flow cytometry we showed that the DNA content is twice that in the parental cell line. Karyotypic examination of metaphasic cells achieved under epifluorescence microscopy revealed a doubled chromosome number in Sf9-HT.

摘要

从鳞翅目昆虫草地贪夜蛾建立的 Sf21 和 Sf9 细胞系在暴露于 37 摄氏度时不会显示热休克蛋白的主要诱导。在 37 摄氏度适应几个月后,我们获得了两个新的细胞系 Sf21-HT 和 Sf9-HT,它们现已在我们实验室建立数年。Sf9-HT 细胞系在 37 摄氏度时的倍增时间比野生型在 28 摄氏度时略短,但细胞致死率导致更早的生长停滞。热适应细胞总脂质提取物的组成显示鞘磷脂与磷脂酰胆碱的比例更高,饱和脂肪酸的百分比更高,这是耐热性所需的较低膜流动性所预期的。Sf9-HT 的细胞体积翻倍,通过流式细胞术我们表明 DNA 含量是亲代细胞系的两倍。在落射荧光显微镜下对中期细胞进行的核型检查显示 Sf9-HT 中的染色体数翻倍。

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