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大规模昆虫和植物细胞培养。

Large-scale insect and plant cell culture.

作者信息

Taticek R A, Lee C W, Shuler M L

机构信息

School of Chemical Engineering, Cornell University, Ithaca, New York 14853-5201.

出版信息

Curr Opin Biotechnol. 1994 Feb;5(2):165-74. doi: 10.1016/s0958-1669(05)80031-x.

Abstract

Currently, insect and plant cell cultures are not widely used to make products of commercial interest, largely because the development of large-scale cultivation methods is still in its infancy. With the advances made over the past year, some of the limitations associated with scale-up of these two types of expression system have been addressed. Increasing the oxygen supply and the concentration of various nutrients supplied to insect cells after infection has enabled high specific protein production to be maintained to higher cell densities than ever before, improving overall volumetric yields. Detailed work has focused on the capacity of insect cells to carry out complex post-translational modifications; however, as yet, evidence is conflicting as to the extent of protein processing and complex glycosylation possible in infected cells. In plant cell culture, the accepted axioms concerning large-scale culture have been re-examined. Recent studies have assessed culture at high cell densities and the constraints in reactor design resulting from the 'shear sensitivity' of plant cells. Results show that, as cell densities increase, alterations occur in the pathways of secondary metabolism, leading to decreases in specific productivity. The use of nutrient supplements and a medium cycling strategy shows promise for increasing and sustaining product formation. Furthermore, the importance of dissolved gas composition has been clearly demonstrated by use of a gas recirculation reactor. Reports of taxol and vindoline production in vitro demonstrate the potential and the necessity for further research in scale-up of plant cell culture.

摘要

目前,昆虫和植物细胞培养尚未广泛用于生产具有商业价值的产品,这主要是因为大规模培养方法的开发仍处于起步阶段。随着过去一年取得的进展,这两种表达系统在放大过程中存在的一些局限性已得到解决。增加感染后供应给昆虫细胞的氧气供应和各种营养物质的浓度,使得在比以往更高的细胞密度下仍能维持高特异性蛋白质产量,从而提高了总体积产量。详细的工作集中在昆虫细胞进行复杂的翻译后修饰的能力上;然而,关于感染细胞中蛋白质加工和复杂糖基化的程度,目前证据相互矛盾。在植物细胞培养中,有关大规模培养的公认原则已被重新审视。最近的研究评估了高细胞密度下的培养以及植物细胞“剪切敏感性”给反应器设计带来的限制。结果表明,随着细胞密度的增加,次生代谢途径会发生变化,导致比生产率下降。使用营养补充剂和培养基循环策略有望提高并维持产物形成。此外,通过使用气体循环反应器,已清楚地证明了溶解气体组成的重要性。体外生产紫杉醇和长春质碱的报道证明了在植物细胞培养放大方面进一步研究的潜力和必要性。

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