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CHO 细胞生长和重组干扰素-γ产生:BSA、普朗尼克和脂质的影响。

CHO cell growth and recombinant interferon-γ production: Effects of BSA, Pluronic and lipids.

机构信息

Research School of Biosciences, Biological Laboratory, University of Kent, CT2 7NJ, Canterbury, UK.

出版信息

Cytotechnology. 1995 Jan;19(1):27-36. doi: 10.1007/BF00749752.

Abstract

The role of bovine serum albumin in mammalian cell cultures and the possibility of its substitution by other components in a serum-free medium has been investigated. In this study, BSA was shown to be important for growth and product formation in CHO cells expressing recombinant human interferon-γ. There were indications that its stimulating growth effect was dependent on the source of BSA used and probably was related to the purification procedure used for the production of the desired albumin fraction. Cell growth did not occur in the absence of BSA but at low concentration (1 mg ml(-1)) it was stimulated by the addition of a combination of a commercial lipid mixture plus Pluronic F68. However, under the latter conditions IFN-γ production was adversely effected. The importance of individual lipid components was investigated using a statistical approach based on a Plackett-Burman design. Linoleic acid was identified as a positive variable for cell growth while cholesterol was identified as a negative variable for both cell growth and IFN-γ production. When a combination of linoleic acid plus Pluronic F68 was included in the formulation of low BSA medium, cell growth was similar to that at high BSA concentration (5 mg ml(-1)) but the IFN-γ concentration was significantly reduced (ca. 45%).

摘要

已研究牛血清白蛋白(BSA)在哺乳动物细胞培养中的作用,以及其在无血清培养基中是否可被其他成分替代的可能性。本研究表明,BSA 对表达重组人干扰素-γ的 CHO 细胞的生长和产物形成很重要。有迹象表明,其促进生长的作用取决于所用 BSA 的来源,可能与所需白蛋白级分的生产所用的纯化程序有关。在没有 BSA 的情况下细胞不会生长,但在低浓度(1mg/ml)下,添加商业脂质混合物加 Pluronic F68 的组合会刺激细胞生长。然而,在后一种情况下 IFN-γ 的产量受到不利影响。使用基于 Plackett-Burman 设计的统计方法研究了单个脂质成分的重要性。亚油酸被确定为细胞生长的正变量,而胆固醇被确定为细胞生长和 IFN-γ 产生的负变量。当在低 BSA 培养基的配方中加入亚油酸加 Pluronic F68 的组合时,细胞生长与高 BSA 浓度(5mg/ml)相似,但 IFN-γ 浓度显著降低(约 45%)。

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