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Title: Transcriptional profiling of batch and fed-batch protein-free 293-HEK cultures
Authors: Lee, Y.Y.
Wong, K.T.K.
Nissom, P.M.
Wong, D.C.F.
Yap, M.G.S. 
Keywords: 293-HEK
Cell culture
Chemically defined
Issue Date: Jan-2007
Citation: Lee, Y.Y., Wong, K.T.K., Nissom, P.M., Wong, D.C.F., Yap, M.G.S. (2007-01). Transcriptional profiling of batch and fed-batch protein-free 293-HEK cultures. Metabolic Engineering 9 (1) : 52-67. ScholarBank@NUS Repository.
Abstract: 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. © 2006.
Source Title: Metabolic Engineering
ISSN: 10967176
DOI: 10.1016/j.ymben.2006.08.006
Appears in Collections:Staff Publications

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