Sample Preparation for Metabolomics: A Method to Prepare Cell Samples for Metabolite Profiling

Abstract

Source: Zhao, Y. C. et. al., Systems Biology of Metabolic Regulation by Estrogen Receptor Signaling in Breast Cancer. J. Vis. Exp. (2016).

This video describes a technique of preparing breast cancer cell samples for metabolite profiling. Breast cancer cells are treated with estradiol, which binds to the estrogen receptor alpha and causes change in the metabolite composition.  

Protocol

1. Metabolomics Assay Sample Preparation Cell Culture and Treatment Seed 400,000 MCF7 cells per 10 cm plate in growth medium. For each treatment prepare three samples. Use two plates in each sample to receive enough cells for the detection. On day 3, remove the medium and add 5 ml fresh medium with or without 10-8 M Estradiol or E2 to the cells. Treat the cells for 24 hr (37 °C in humidified 5% CO2 atmosphere). Sample Collection …