文献类型：期刊文献

英文题名：Improvement of Vero cell growth in glutamate-based culture by supplementing ammoniagenic compounds

作者：Huang, Haiyan[1]; Yi, Xiaoping[1]; Zhang, Yuanxing[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2006

卷号：41

期号：12

起止页码：2386

外文期刊名：PROCESS BIOCHEMISTRY

收录：;EI(收录号：20064310200114);WOS:【SCI-EXPANDED(收录号:WOS:000242259100005)】；

语种：英文

外文关键词：alanine; asparagine; glutamate; glutamine-free; glutamine synthetase; Vero cell

摘要：Ammonia has been identified as one of the most inhibitory substances for mammalian cells. We have attempted to develop a less-ammoniagenic medium for the growth of Vero cells by substitution of glutamine with glutamate. In spite of reduced ammonia formation, Vero cells cultured in glutamate-based medium (DMEM-glu) could not grow normally as in glutamine-based medium (DMEM-gln). After Vero cells adapted to DMEM-glu, alanine was consumed instead of accumulated and both asparagine and glutamine were almost undetectable, indicating the lacking for aminonitrogen. By supplementing NH4Cl, the growth was significantly improved and the cellular uptake of glutamate from medium was greatly increased. However the growth was still not restored to the level in DMEM-gln, likely due to ammonia toxicity. Asparagine was chosen to support the growth of Vero cells in DMEM-glu, formulating DMEM-glu-asn. This replacement reduced ammonia formation by 79% and increased cell yields by 34% compared with DMEM-gln. After Vero cells adapted to DMEM-glu-asn, glutamine synthetase (GS) activity was elevated by 3.8 folds compared with control in DMEM-gln. In DMEM-glu-asn Vero cell growth was arrested by the specific GS inhibitor, methionine sulphoximine. This arrest affirmed the essential role of GS in glutamine synthesis and disconfirmed the potential role of asparagine synthase (AS) in glutamine formulation (also asparagine utilization). (c) 2006 Elsevier Ltd. All rights reserved.