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无血清培养条件下烟青虫细胞系的生长、病毒生产和代谢。

Growth, viral production and metabolism of a Helicoverpa zea cell line in serum-free culture.

机构信息

Department of Chemical Engineering, The University of Queensland, Brisbane, Queensland, 4072, Australia(e-mail,

出版信息

Cytotechnology. 2003 Sep;42(3):109-20. doi: 10.1023/B:CYTO.0000015795.46813.44.

Abstract

Insect cell cultures have been extensively utilised for means of production for heterologous proteins and biopesticides. Spodoptera frugiperda (Sf9) and Trichoplusia ni (High Fivetrade mark) cell lines have been widely used for the production of recombinant proteins, thus metabolism of these cell lines have been investigated thoroughly over recent years. The Helicoverpa zea cell line has potential use for the production of a biopesticide, specifically the Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HaSNPV). The growth, virus production, nutrient consumption and waste production of this cell line was investigated under serum-free culture conditions, using SF900II and a low cost medium prototype (LCM). The cell growth (growth rates and population doubling time) was comparable in SF900II and LCM, however, lower biomass and cell specific virus yields were obtained in LCM. H. zea cells showed a preference for asparagine over glutamine, similar to the High Fivetrade mark cells. Ammonia was accumulated to significantly high levels (16 mM) in SF900II, which is an asparagine and glutamine rich medium. However, given the absence of asparagine and glutamine in the medium (LCM), H. zea cells adapted and grew well in the absence of these substrates and no accumulation of ammonia was observed. The adverse effect of ammonia on H. zea cells is unknown since good production of biologically active HaSNPV was achieved in the presence of high ammonia levels. H. zea cells showed a preference for maltose even given an abundance supply of free glucose. Accumulation of lactate was observed in H. zea cell cultures.

摘要

昆虫细胞培养已被广泛用于生产异源蛋白和生物农药。草地贪夜蛾(Sf9)和三化螟(High Five 商标)细胞系已被广泛用于生产重组蛋白,因此近年来对这些细胞系的代谢进行了深入研究。玉米螟细胞系具有生产生物农药的潜力,特别是玉米螟核型多角体病毒(HaSNPV)。在无血清培养条件下,使用 SF900II 和低成本培养基原型(LCM),研究了该细胞系的生长、病毒生产、营养消耗和废物产生情况。SF900II 和 LCM 中的细胞生长(生长速率和倍增时间)相当,但 LCM 中的生物量和细胞特异性病毒产量较低。玉米螟细胞对天冬酰胺的偏好超过谷氨酰胺,与 High Five 细胞相似。在 SF900II 中,氨积累到了非常高的水平(16mM),这是一种富含天冬酰胺和谷氨酰胺的培养基。然而,由于培养基(LCM)中没有天冬酰胺和谷氨酰胺,玉米螟细胞适应并在没有这些底物的情况下生长良好,没有观察到氨的积累。高浓度氨对玉米螟细胞的不利影响尚不清楚,因为在高氨浓度下仍能产生具有生物活性的 HaSNPV。玉米螟细胞即使有大量游离葡萄糖供应,也偏爱麦芽糖。在玉米螟细胞培养物中观察到乳酸盐的积累。

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