Mammalian cell cultures as models for metabolomic studies / Nel Z.

• Main Author: Nel, Maria Elizabeth.
• Published: North-West University 2013
• Online Access: http://hdl.handle.net/10394/8205

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.