Cell culture metabolomics: applications and future directions

Čuperlović-Culf, Miroslava; Barnett, David A.; Culf, Adrian S.; Chute, Ian C.

doi:10.1016/j.drudis.2010.06.012

Cited by 190 publications

(158 citation statements)

References 76 publications

Supporting

Mentioning

152

Contrasting

Unclassified

Order By: Relevance

“…This is particularly an issue is studies involving comparative analysis of several different cell types, all of which might contain different levels of glucose, glutamine and lactate, as well as other nutrients and additives, which will probably lead to differences in the metabolome of the cells. If possible, it is recommended to use the same growth medium for all cell lines in the study to reduce variance in metabolic profile that can be caused by the medium (Cuperlovic-Culf et al, 2010). The standard enhancement of cell culture medium with serum of animal origin can add another level of complexity in cell growth condition optimization.…”

Section: Cell Culture Growth Stimulationmentioning

confidence: 99%

“…Metabolic footprinting is technically simple because it requires only centrifugation to separate culture media and cells before the analysis. Metabolic fingerprinting, although much more technically challenging because it requires metabolite extraction from cells, provides more complete information about cellular metabolic processes (Cuperlovic-Culf et al, 2010). Recently a study related with different metabolite extraction protocols for mammalian cell culture was published (Dietmair et al, 2010).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Metabolomics and Mammalian Cell Culture

Luz-Hdez¹

2012

Metabolomics

View full text Add to dashboard Cite

Section: Cell Culture Growth Stimulationmentioning

confidence: 99%

mentioning

confidence: 99%

Metabolomics and Mammalian Cell Culture

Luz-Hdez¹

2012

Metabolomics

View full text Add to dashboard Cite

“…Metabolomics, therefore, is considered a more approachable methodology than genomics or proteomics, particularly in translating results to phenotypic changes. Because metabolic changes represent the final outcome of all physiological processes, metabolomics is a relevant approach to study toxicity (Robertson, 2005 In vitro toxicity studies using metabolomics have been rather limited so far (Cuperlovic-Culf et al, 2010). However, in vitro applications are likely to increase due to some advantages over the analysis of body fluids.…”

Section: Metabolomicsmentioning

confidence: 99%

“…Still, a limitation of the MS-based approach is that often not all metabolites can be identified. The data analysis procedures of NMR and MS datasets depend upon the metabolomics study performed (Cuperlovic-Culf et al, 2010). A profiling study is a hypothesis-driven approach based on pre-selected metabolites relevant for a specific pathway, disease, or toxic perturbation.…”

Section: The Potential Role Of Omics Technologies In the Visionmentioning

confidence: 99%

Current standing and future prospects for the technologies proposed to transform toxicity testing in the 21st century

Vliet

2011

ALTEX

View full text Add to dashboard Cite

“…This is especially the case when nutrient levels in feeds are progressively increased for high cell density infections, leading to perturbations such as osmotic shock and toxin accumulation [7]. A systems biology approach, involving the integration of various omics platforms such as transcriptomics and metabolomics into genome-scale computational models, could bring about better elucidation of complex biological systems, and offer rational strategies towards bioprocess optimization [8,9]. With this in mind, a methodology was developed in this study for quantitative intracellular metabolite analysis of insect cells, to enable the application of metabolomics for the baculovirus-insect cell expression system.…”

Section: Introductionmentioning

confidence: 99%

Metabolomic study of baculovirus infected insect cells to identify metabolite based strategies for improving virus yields

Tran¹

View full text Add to dashboard Cite

The baculovirus insect cell system has been used widely for biopesticide applications and for the production of recombinant proteins, vaccines and vectors for gene delivery. To be commercially viable, especially for the low margin applications of biopesticides and veterinary vaccines, the yield of this production system needs to be improved. One of the important approaches is the infection of insect cell cultures at high cell densities in order to maximize volumetric production. However, the cell specific yield gradually reduces with increasing infection cell density (ICD). As a result, the volumetric production can not improve further regardless of efforts to increase the uninfected cell density in batch or fedbatch cultures. The phenomenon known as "the cell density effect" appears not only during the late stage of protein production, but also during the early stage of virus replication and transcription. Discovering the changes in intra and extracellular metabolites between low and high ICDs may reveal evidence for improving yields. This PhD thesis focuses on the development of an intracellular metabolite extraction protocol for infected insect cell cultures and for measuring intra and extracellular metabolites of infected cells of different baculovirus infected insect cell systems.Prior to analyzing the intracellular metabolites of low and high ICDs, an extraction protocol was developed and validated for insect cell cultures, especially infected insect cells. The ice-cold saline quenching solution developed for mammalian cells did not work well for insect cells, especially for infected cultures. Therefore, a modification of the quenching solution was made by an addition of 0.2% Pluronic® F-68 that successfully protected the cell membrane during quenching and washing procedures. In addition, one wash was found to be enough for removal of medium originated metabolites adhering on the surface of cell membranes. The ATP recovery (over the direct extract) of the extraction protocol using ice-cold saline quenching solution plus 0.2% Pluronic® F-68 after one wash was almost 90% for Spodoptera frugiperda (Sf9) cells infected with a recombinant Autographa californica multiple nucleopolyhedrovirus (rAcMNPV) and Helicoverpa zea (HzAM1) cells infected with a wild-type Helicoverpa armigera single nucleopolyhedrovirus (HearNPV) at 24 hours post infection (hpi).The intracellular metabolite patterns and substrate consumption rates of low versus high ICDs was first investigated for the Sf9/rAcMNPV system in Sf900™III medium. A preliminary study was conducted to identify how late post-infection the cell membrane integrity was maintained during the quenching and washing processes. The result showed that the extraction protocol is efficient up to 48 hpi for infected Sf9 cells. Levels of intracellular nucleotides (tri-phosphates), amino acids (AA), and TCA cycle intermediates iii (α-ketoglutarate, pyruvate) during an infection at a low ICD (2×10 6 cells/mL) were shown to be higher than those during an infection at a high...

show abstract

Cell culture metabolomics: applications and future directions

Cited by 190 publications

References 76 publications

Metabolomics and Mammalian Cell Culture

Metabolomics and Mammalian Cell Culture

Current standing and future prospects for the technologies proposed to transform toxicity testing in the 21st century

Metabolomic study of baculovirus infected insect cells to identify metabolite based strategies for improving virus yields

Contact Info

Product

Resources

About