Abstract
Use of solid phase microextraction (SPME) for cell culture metabolomic analysis allows for the attainment of more sophisticated data from in vitro cell cultures. Moreover, considering that SPME allows the implementation of multiple extractions from the same sample due to its non/low-depletive nature, time course studies using the same set of samples are thus facilitated via this method. Such an approach results in a reduction in the number of samples needed for analysis thus eliminates inter-batch variability related to biological variation occurring during cell culturing. The current work aims to demonstrate the capability of SPME for measurements of combretastatin A4 (CA4) effectiveness on non-small cell cancer cell line. A cultivation protocol was established in the 96-well plate, and a fiber format of SPME was selected for metabolite extraction. The extracellular metabolic pattern of cells was changed after administration of the tested drug. This suggests pharmacological activity of the administered compound towards the studied cell line model. Results support that the use of direct immersion SPME for analysis of cell cultures does not affect cells growth or contaminate sample. Consequently, SPME allows the attainment of accurate information regarding drug uptake, metabolism, and metabolomic changes in the studied cells induced by exposure to the drug simultaneously in a single experiment.
Highlights
Nowadays, most standard anticancer therapies incorporate the use of cytotoxic agents
Different drug dosing might result in different metabolome changes. Such comparison was beyond the scope of this study, which main goal was to demonstrate the applicability of solid phase microextraction (SPME) technology to cell line studies in different experimental setups
Solid phase microextraction is presented in the current work as a suitable tool for high throughput tracking of metabolic changes in in vitro systems, either as part of one time-point investigations, or in time course studies, in which case SPME offers the possibility of multiple analyses from the same samples due to its minimally-invasive nature, and negligible depletion capabilities
Summary
Most standard anticancer therapies incorporate the use of cytotoxic agents Due to their lack of selectivity to cancer tissue, their administration often causes wide-ranging side effects[1]. Metabolomics studies can reveal vital information regarding small molecules within a biological sample, enabling a better understanding of the particular biochemistry of a given matrix, such as that of cancer cells In this respect, solid phase microextraction (SPME) is a valuable platform for metabolomics studies, having been utilized for investigations pertaining to a wide variety of biological matrices such as fruits, leaves, animal tissues (both in and ex vivo), and biofluids (serum, plasma, whole blood, urine saliva), among others[12]. As we employed two different setups of SPME, the flexibility of the extraction method was shown as both SPME protocols enable performing other assays on cells
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