Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture

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, t...

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Autores principales:	Andrea Antonio Ellero, Iman van den Bout, Maré Vlok, Allan Duncan Cromarty, Tracey Hurrell
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/32eca0f8407849fba20442dca1f15db0
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spelling	oai:doaj.org-article:32eca0f8407849fba20442dca1f15db02021-12-02T15:00:13ZContinual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture10.1038/s41598-021-89907-92045-2322https://doaj.org/article/32eca0f8407849fba20442dca1f15db02021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89907-9https://doaj.org/toc/2045-2322Abstract 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.Andrea Antonio ElleroIman van den BoutMaré VlokAllan Duncan CromartyTracey HurrellNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Andrea Antonio Ellero Iman van den Bout Maré Vlok Allan Duncan Cromarty Tracey Hurrell Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture
description	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.
format	article
author	Andrea Antonio Ellero Iman van den Bout Maré Vlok Allan Duncan Cromarty Tracey Hurrell
author_facet	Andrea Antonio Ellero Iman van den Bout Maré Vlok Allan Duncan Cromarty Tracey Hurrell
author_sort	Andrea Antonio Ellero
title	Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture
title_short	Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture
title_full	Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture
title_fullStr	Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture
title_full_unstemmed	Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture
title_sort	continual proteomic divergence of hepg2 cells as a consequence of long-term spheroid culture
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/32eca0f8407849fba20442dca1f15db0
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Continual proteomic divergence of HepG2 cells as a consequence of long-term spheroid culture

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