کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
31951 | 44858 | 2007 | 16 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Transcriptional profiling of batch and fed-batch protein-free 293-HEK cultures Transcriptional profiling of batch and fed-batch protein-free 293-HEK cultures](/preview/png/31951.png)
Dynamic nutrient feeding to control glutamine at low levels in protein-free fed-batch cultures of 293-human embryonic kidney (HEK) cells achieved cell concentrations of 6×106 cells/ml. This represented a 4-fold improvement in cell concentration compared to batch cultures. Reduction in glutamine and glucose consumption, as well as lactate and ammonia production, were also observed in these fed-batch cultures. High virus production titers of 3×1011 pfu/ml were achieved in fed-batch cultures which were 10,000-fold higher than batch cultures.An investigation of the transcriptional regulation of the metabolic changes associated with the batch and the low-glutamine fed-batch cultures using DNA microarray was conducted. This analysis provides better understanding of the transcriptional regulatory mechanism resulting in the observed physiological changes. Transcriptional profiling of cells from the mid-exponential, late exponential and stationary phases of both the batch and fed-batch were undertaken using an 18,000 element human chip. Transcriptional profiles were ontologically classified to provide a global view of the genetic changes. Furthermore, a pathway-oriented analysis focusing on cellular metabolism was conducted to reveal the dynamic regulation of genes related to amino acid metabolism, tRNA synthetases, TCA cycle, electron transport chain and glycolysis.
Journal: Metabolic Engineering - Volume 9, Issue 1, January 2007, Pages 52–67