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Characterization and reproducibility of HepG2 hanging drop spheroids toxicology in vitro. | LitMetric

Characterization and reproducibility of HepG2 hanging drop spheroids toxicology in vitro.

Toxicol In Vitro

Department of Pharmacology, Faculty of Health Sciences, School of Medicine, University of Pretoria, Private Bag X323, Arcadia, 0007, South Africa. Electronic address:

Published: August 2018

AI Article Synopsis

  • The study explores using 3D spheroid cultures of HepG2 cells, particularly through a high-throughput hanging drop technique, to achieve more accurate liver cell characteristics for testing.
  • Findings revealed variations in cell behavior based on factors such as cell stocks, seeding density, culture medium volume, and matrix additives, which raises concerns about the consistency and reliability of these cultures in modeling liver responses.

Article Abstract

Hepatotoxicity remains a major challenge in drug development despite preclinical toxicity screening using hepatocytes of human origin. To overcome some limitations of reproducing the hepatic phenotype, more structurally and functionally authentic cultures in vitro can be introduced by growing cells in 3D spheroid cultures. Characterisation and reproducibility of HepG2 spheroid cultures using a high-throughput hanging drop technique was performed and features contributing to potential phenotypic variation highlighted. Cultured HepG2 cells were seeded into Perfecta 3D® 96-well hanging drop plates and assessed over time for morphology, viability, cell cycle distribution, protein content and protein-mass profiles. Divergent aspects which were assessed included cell stocks, seeding density, volume of culture medium and use of extracellular matrix additives. Hanging drops are advantageous due to no complex culture matrix being present, enabling background free extractions for downstream experimentation. Varying characteristics were observed across cell stocks and batches, seeding density, culture medium volume and extracellular matrix when using immortalized HepG2 cells. These factors contribute to wide-ranging cellular responses and highlights concerns with respect to generating a reproducible phenotype in HepG2 hanging drop spheroids.

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Source
http://dx.doi.org/10.1016/j.tiv.2018.02.013DOI Listing

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