Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture
Date
2021
Journal Title
Journal ISSN
Volume Title
Publisher
Nature
Abstract
Three-dimensional models are considered a powerful tool for improving the concordance between in vitro and in vivo phenotypes. However, the duration of spheroid culture may influence the degree of correlation between these counterparts. When using immortalised cell lines as model systems, the assumption for consistency and reproducibility is often made without adequate characterization or validation. It is therefore essential to define the biology of each spheroid model by investigating proteomic dynamics, which may be altered relative to culture duration. As an example, we assessed the influence of culture duration on the relative proteome abundance of HepG2 cells cultured as spheroids, which are routinely used to model aspects of the liver. Quantitative proteomic profiling of whole cell lysates labelled with tandem-mass tags was conducted using liquid chromatography-tandem mass spectrometry (LC–MS/MS). In excess of 4800 proteins were confidently identified, which were shared across three consecutive time points over 28 days. The HepG2 spheroid proteome was divergent from the monolayer proteome after 14 days in culture and continued to change over the successive culture time points. Proteins representing the recognised core hepatic proteome, cell junction, extracellular matrix, and cell adhesion proteins were found to be continually modulated.
Description
CITATION: Ellero, A. A. et al. 2021. Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture. Scientific Reports, 11:10917, doi:10.1038/s41598-021-89907-9.
The original publication is available at https://www.nature.com
The original publication is available at https://www.nature.com
Keywords
HepG2 cells, Proteomics -- Dynamics, Phenotype, Liver cells -- Effect of drugs on, Spheroids
Citation
Ellero, A. A. et al. 2021. Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture. Scientific Reports, 11:10917, doi:10.1038/s41598-021-89907-9.