Dynamics of Amino Acid Metabolism, Gene Expression, and Circulomics in a Recombinant Chinese Hamster Ovary Cell Line Adapted to Moderate and High Levels of Extracellular Lactate.
Dynamics of Amino Acid Metabolism, Gene Expression, and Circulomics in a Recombinant Chinese Hamster Ovary Cell Line Adapted to Moderate and High Levels of Extracellular Lactate.
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DOI：
10.3390/genes14081576
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发表时间：
2023-08-02
期刊：
影响因子：
3.5
通讯作者：
Saski, Christopher A.
中科院分区：
文献类型：
--
作者：
Chitwood, Dylan G.;Uy, Lisa;Fu, Wanfang;Klaubert, Stephanie R.;Harcum, Sarah W.;Saski, Christopher A.
细胞培养物中代谢废物的积累可降低产品质量、降低生产率并引发细胞凋亡。已尝试通过多种工艺和基因工程途径限制或去除中国仓鼠卵巢（CHO）细胞培养物中的非预期废物，并取得了不同程度的成功。一项研究证明了一种简单的方法，以减少乳酸和氨的生产在CHO细胞与适应细胞外乳酸;然而，适应背后的机制是不确定的。为了解决这一深刻的差距，本研究表征了重组CHO K-1细胞系的表型，该细胞系逐渐适应中等和高水平的细胞外乳酸，并研究了染色体外环状DNA（eccDNA）和基因表达对适应过程的基因组含量和作用。在eccDNA上观察到超过500个基因。值得注意的是，超过1000个基因被观察到在不同水平的乳酸适应差异表达，而只有137个基因被发现在不适应的细胞和细胞之间的差异表达适应生长在高水平的乳酸;这表明随机开关作为一个潜在的压力适应机制在CHO细胞。此外，这些数据表明丙氨酸生物合成是CHO细胞中过量乳酸的潜在应激缓解机制。
The accumulation of metabolic wastes in cell cultures can diminish product quality, reduce productivity, and trigger apoptosis. The limitation or removal of unintended waste products from Chinese hamster ovary (CHO) cell cultures has been attempted through multiple process and genetic engineering avenues with varied levels of success. One study demonstrated a simple method to reduce lactate and ammonia production in CHO cells with adaptation to extracellular lactate; however, the mechanism behind adaptation was not certain. To address this profound gap, this study characterizes the phenotype of a recombinant CHO K-1 cell line that was gradually adapted to moderate and high levels of extracellular lactate and examines the genomic content and role of extrachromosomal circular DNA (eccDNA) and gene expression on the adaptation process. More than 500 genes were observed on eccDNAs. Notably, more than 1000 genes were observed to be differentially expressed at different levels of lactate adaptation, while only 137 genes were found to be differentially expressed between unadapted cells and cells adapted to grow in high levels of lactate; this suggests stochastic switching as a potential stress adaptation mechanism in CHO cells. Further, these data suggest alanine biosynthesis as a potential stress-mitigation mechanism for excess lactate in CHO cells.
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影响因子：
2.9
作者：
Altamirano, C;Paredes, C;Gòdia, F
通讯作者：
Gòdia, F
DOI：
10.1007/bf00369401
发表时间：
1993-10-01
期刊：
BIOPROCESS ENGINEERING
影响因子：
--
作者：
CONVERTI, A;SOMMARIVA, C;FERRAIOLO, G
通讯作者：
FERRAIOLO, G
DOI：
10.1093/bioinformatics/bts565
发表时间：
2012-12-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Fu L;Niu B;Zhu Z;Wu S;Li W
通讯作者：
Li W
影响因子：
3.5
作者：
de Sury, R;Martinez, P;Issartel, JP
通讯作者：
Issartel, JP
影响因子：
7
作者：
Cohen, S;Yacobi, K;Segal, D
通讯作者：
Segal, D