| Summary: | The use of cultured cells in metabolomic studies is receiving more and more
attention. There are many advantages when using cultured cells in metabolomic
studies, for example cultured cells can easily be manipulated for the purpose of the
experiment. This creates many opportunities for metabolomics studies, for example
cell cultures can offer an alternative manner of drug testing. Even though the use of
cultured cells in metabolomic studies is very promising and they create many
opportunities for metabolomic research, there are still challenges that create
obstacles in this research. One of the challenges is that present analytical
technologies do not always fully meet the requirements for metabolomics. There is,
however, much effort going into optimising the methods concerning cultured cells
and metabolomics, but there is a lack of attention when it comes to the sample
preparation which is initiated by quenching. The aim of this study was to investigate
cultured cells as models for metabolomics investigations and to standardise a proper
quenching method for a metabolomics analysis of mammalian cultured cells.
A quenching method adapted from the literature was evaluated for the cell line used
in this study, namely HeLa. Metabolites of the central carbon metabolism were
targeted, using a published list. This method was tested for its effectiveness by
introducing the samples to waiting periods (0, 3, 6 and 24 hours) before extraction
after immediate quenching. Results indicated that the entire metabolism under study
was not effectively quenched. The optimum composition and temperature for this
quenching method were also investigated by comparing three different quenching
methods derived from the literature. The results were contradicting. Cell cultures
were exposed to two perturbations (environmental and genetic) to investigate if
these perturbations can be captured and measured by using metabolomics as an
instrument. There was a significant difference between control groups and the
groups exposed to the different perturbations. The results gained from this study
indicate that it is definitely possible to use cultured cells in metabolomics studies. === Thesis (MSc (Biochemistry))--North-West University, Potchefstroom Campus, 2012.
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