Use of a non-hepatic cell line highlights limitations associated with cell-based assessment of metabolically induced toxicity

Weyers, Carli; Dingle, Laura Margaret Kirkpatrick; Wilhelmi, Brendan; Edkins, Adrienne L.; Veale, Clinton G. L.

doi:10.1080/01480545.2019.1585869

Cited by 2 publications

(2 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The impact of a dynamic baseline would be evident in experiments such as drug testing for efficacy or toxicity, if cells which rapidly form spheroids, were cultured for 7, 10, or 28 days. While recognised as limited in their biotransformation capacity 20 , HepG2 cell spheroids are used to investigate genotoxicity 21 , and predict hepatotoxicity 22 , 23 . Phenotypic transitions as a result of growth in 3D have been investigated at the gene transcription level 24 which while informative, does not account for the fact that only 20–50% of transcribed genes result in expression of functional proteins 25 , which would limit the perceived utility of these model systems.…”

Section: Introductionmentioning

confidence: 99%

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

Ellero

Bout

Vlok

et al. 2021

Sci Rep

View full text Add to dashboard Cite

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.

show abstract

Section: Introductionmentioning

confidence: 99%

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

Ellero

Bout

Vlok

et al. 2021

Sci Rep

View full text Add to dashboard Cite

show abstract

“…The impact of a dynamic baseline would be evident in experiments such as drug testing for efficacy or toxicity if cells which rapidly form spheroids were first cultured for 7, 10, or 28 days. While recognised as limited in their biotransformation capacity [223], HepG2 cell spheroids are used as a liver model to investigate genotoxicity [224], and predict hepatotoxicity [225,226]. Phenotypic transitions as a result of growth in 3D have been investigated at the gene transcription level [227] which while informative, does not account for the fact that only 20-50% of transcribed genes result in functional expressed proteins [228], and could cast doubt on the perceived utility of model systems.…”

Section: Introductionmentioning

confidence: 99%

ProteomeXchange dataset

View full text Add to dashboard Cite

show abstract

Use of a non-hepatic cell line highlights limitations associated with cell-based assessment of metabolically induced toxicity

Cited by 2 publications

References 36 publications

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

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

ProteomeXchange dataset

Contact Info

Product

Resources

About