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悬浮培养中 Anticarsia gemmatalis 细胞系的生长、代谢和杆状病毒生产。

Growth, metabolism and baculovirus production in suspension cultures of an Anticarsia gemmatalis cell line.

机构信息

Laboratorio de Virología, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, CC 242, S3000ZAA, Santa Fe, República Argentina.

出版信息

Cytotechnology. 2006 Oct;52(2):113-24. doi: 10.1007/s10616-006-9042-3. Epub 2007 Jan 23.

Abstract

The UFL-AG-286 cell line, established from embryonic tissue of the lepidopteran insect Anticarsia gemmatalis, has been identified as a good candidate to be used as a cellular substrate in the development of a process for in vitro production of the Anticarsia gemmatalis multicapsid nucleopolyhedrovirus, a baculovirus widely used as bioinsecticide. In order to characterize the technological properties of this cell line and evaluate its feasibility to use it for the large-scale production of Anticarsia gemmatalis multicapsid nucleopolyhedrovirus, UFL-AG-286 cells were adapted to grow as agitated suspension cultures in spinner-flasks. Batch suspension cultures of adapted cells in serum-supplemented TC-100 medium grew with a doubling time of about 29 h and reached a maximum cell density higher than 3.5 x 10(6) viable cells ml(-1). At the end of the growth period glucose was completely depleted from the culture medium, but L: -lactate was not produced. Amino acids, with the exception of glutamine, were only negligibly consumed or produced. In contrast to other insect cell lines, UFL-AG-286 cells appeared to be unable to synthesize alanine as a metabolic way to dispose the by-product ammonia. The synchronous infection of suspension cultures with Anticarsia gemmatalis multicapsid nucleopolyhedrovirus in the early to medium exponential growth phase yielded high amounts of both viral progenies per cell and reduced the specific demands of UFL-AG-286 cells for the main nutrients.

摘要

UFL-AG-286 细胞系是从鳞翅目昆虫 Anticarsia gemmatalis 的胚胎组织中建立的,已被确定为一种很好的候选细胞基质,可用于开发体外生产 Anticarsia gemmatalis 多壳核多角体杆状病毒的方法,该杆状病毒广泛用作生物杀虫剂。为了表征该细胞系的技术特性,并评估其用于大规模生产 Anticarsia gemmatalis 多壳核多角体杆状病毒的可行性,将 UFL-AG-286 细胞适应在摇瓶中的搅拌悬浮培养中生长。在含有血清的 TC-100 培养基中适应的细胞的分批悬浮培养以约 29 小时的倍增时间生长,并达到超过 3.5 x 10(6)个活细胞/ml(-1)的最大细胞密度。在生长周期结束时,培养基中的葡萄糖被完全耗尽,但未产生 L: - 乳酸。除了谷氨酰胺外,氨基酸的消耗或产生可以忽略不计。与其他昆虫细胞系不同,UFL-AG-286 细胞似乎无法合成丙氨酸作为处理副产物氨的代谢途径。在早期到中指数生长阶段,悬浮培养物的同步感染 Anticarsia gemmatalis 多壳核多角体杆状病毒产生了大量的每细胞病毒产物,并降低了 UFL-AG-286 细胞对主要营养物质的特定需求。

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