Influence of washing and quenching in profiling the metabolome of adherent mammalian cells: a case study with the metastatic breast cancer cell line MDA-MB-231

The Analyst
Rahul Vijay KapooreSeetharaman Vaidyanathan

Abstract

Metabolome characterisation is a powerful tool in oncology. To obtain a valid description of the intracellular metabolome, two of the preparatory steps are crucial, namely washing and quenching. Washing must effectively remove the extracellular media components and quenching should stop the metabolic activities within the cell, without altering the membrane integrity of the cell. Therefore, it is important to evaluate the efficiency of the washing and quenching solvents. In this study, we employed two previously optimised protocols for simultaneous quenching and extraction, and investigated the effects of a number of washing steps/solvents and quenching solvent additives, on metabolite leakage from the adherent metastatic breast cancer cell line MDA-MB-231. We explored five washing protocols and five quenching protocols (including a control for each), and assessed for effectiveness by detecting ATP in the medium and cell morphology changes through scanning electron microscopy (SEM) analyses. Furthermore, we studied the overall recovery of eleven different metabolite classes using the GC-MS technique and compared the results with those obtained from the ATP assay and SEM analysis. Our data demonstrate that a single washing step ...Continue Reading

References

Sep 2, 2003·Biochimica Et Biophysica Acta·Erik M Schwiebert, Akos Zsembery
Apr 7, 2007·Analytical Chemistry·Christoph J BoltenChristoph Wittmann
May 12, 2007·Biotechnology and Applied Biochemistry·Soo Hean Gary Khoo, Mohamed Al-Rubeai
Apr 30, 2008·Plant Methods·Do Yup Lee, Oliver Fiehn
Dec 17, 2008·Analytical Biochemistry·Hilal Taymaz-NikerelWalter M van Gulik
Nov 11, 2009·Annals of Oncology : Official Journal of the European Society for Medical Oncology·O GluzN Harbeck
May 4, 2010·Analytical Biochemistry·Stefanie DietmairJens O Krömer
Jun 12, 2010·Molecular Oncology·Franca PodoUNKNOWN FEMME Consortium
Jul 6, 2010·Drug Discovery Today·Miroslava Cuperlović-CulfIan Chute
Jun 22, 2011·Applied Microbiology and Biotechnology·Friederike SchädelEzequiel Franco-Lara
Jul 19, 2011·Journal of Microbiological Methods·Renata MarcinowskaIzabella Surowiec
Oct 4, 2012·Annals of Oncology : Official Journal of the European Society for Medical Oncology·C CriscitielloC Sotiriou
Sep 2, 2014·Metabolomics : Official Journal of the Metabolomic Society·Preeti PurwahaJohn N Weinstein
Jan 24, 2015·Analytical Biochemistry·Zheng SerJason W Locasale
Oct 9, 2015·Analytical and Bioanalytical Chemistry·Blandine Madji HounoumSylvie Mavel
Sep 21, 2016·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Rahul Vijay Kapoore, Seetharaman Vaidyanathan

❮ Previous
Next ❯

Citations

Apr 15, 2020·Analytical and Bioanalytical Chemistry·Paula Cuevas-DelgadoCoral Barbas
Sep 16, 2020·Analytical Methods : Advancing Methods and Applications·Yun LuoHaijun Zhang
Jul 3, 2017·Cellular and Molecular Life Sciences : CMLS·Sarah HaytonRobert D Trengove
Sep 24, 2020·International Journal of Molecular Sciences·Aneta BalcerczykGilles J P Rautureau
May 28, 2019·Metabolomics : Official Journal of the Metabolomic Society·Jordan WilkinsEugenia Trushina
Jun 20, 2020·Analytical and Bioanalytical Chemistry·Manhal MiliBénédicte Elena-Herrmann
Oct 22, 2020·BMC Complementary Medicine and Therapies·Tanaporn HengpratomGriangsak Eumkeb
Nov 11, 2019·Journal of Pharmaceutical and Biomedical Analysis·Malena ManziMaría Eugenia Monge
Mar 9, 2021·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jinfeng ZhuHongmin Liu
May 14, 2021·Critical Reviews in Biotechnology·Rahul Vijay KapooreSeetharaman Vaidyanathan

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.