title: ¹H NMR spectroscopy profiling of metabolic reprogramming of Chinese hamster ovary cells upon a temperature shift during culture.
abstract: We report an NMR based approach to determine the metabolic reprogramming of Chinese hamster ovary cells upon a temperature shift during culture by investigating the extracellular cell culture media and intracellular metabolome of CHOK1 and CHO-S cells during culture and in response to cold-shock and subsequent recovery from hypothermic culturing. A total of 24 components were identified for CHOK1 and 29 components identified for CHO-S cell systems including the observation that CHO-S media contains 5.6 times the level of glucose of CHOK1 media at time zero. We confirm that an NMR metabolic approach provides quantitative analysis of components such as glucose and alanine with both cell lines responding in a similar manner and comparable to previously reported data. However, analysis of lactate confirms a differentiation between CHOK1 and CHO-S and that reprogramming of metabolism in response to temperature was cell line specific. The significance of our results is presented using principal component analysis (PCA) that confirms changes in metabolite profile in response to temperature and recovery. Ultimately, our approach demonstrates the capability of NMR providing real-time analysis to detect reprogramming of metabolism upon cellular perception of cold-shock/sub-physiological temperatures. This has the potential to allow manipulation of metabolites in culture supernatant to improve growth or productivity.
authors.affiliation: Centre for Molecular Processing and School of Biosciences, University of Kent, Canterbury, Kent, United Kingdom.
authors.forename: Jane L
authors.initials: JL
authors.lastname: Wagstaff
authors.forename: Rosalyn J
authors.initials: RJ
authors.lastname: Masterton
authors.forename: Jane F
authors.initials: JF
authors.lastname: Povey
authors.forename: C Mark
authors.initials: CM
authors.lastname: Smales
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Howard
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D005110:Extracellular Space / Q000378:metabolism; D005260:Female; D005947:Glucose / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D055442:Metabolome; D010053:Ovary / Q000166:cytology*; D013696:Temperature
chemical_list: D000596:Amino Acids; D019344:Lactic Acid; D005947:Glucose