Characterization and reproducibility of HepG2 hanging drop spheroids toxicology in vitro

Show simple item record Hurrell, Tracey Ellero, Andrea Antonio Masso, Zelie Flavienne Cromarty, Allan Duncan 2018-08-07T10:02:06Z 2018-08
dc.description.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. en_ZA
dc.description.department Pharmacology en_ZA
dc.description.embargo 2019-08-01
dc.description.librarian hj2018 en_ZA
dc.description.sponsorship A National Research Foundation of South Africa Thuthuka grant (Grant No. 87880). en_ZA
dc.description.uri en_ZA
dc.identifier.citation Hurrell, T., Ellero, A.A., Masso, Z.F. & Cromarty, A.D. 2018, 'Characterization and reproducibility of HepG2 hanging drop spheroids toxicology in vitro', Toxicology in Vitro, vol. 50, pp. 86-94. en_ZA
dc.identifier.issn 0887-2333 (print)
dc.identifier.issn 1879-3177 (online)
dc.identifier.other 10.1016/j.tiv.2018.02.013
dc.language.iso en en_ZA
dc.publisher Elsevier en_ZA
dc.rights © 2018 Elsevier Inc. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Toxicology in Vitro. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. A definitive version was subsequently published in Toxicology in Vitro, vol. 50, pp. 86-94, 2018. doi : 10.1016/j.tiv.2018.02.013. en_ZA
dc.subject HepG2 cells en_ZA
dc.subject Hepatotoxicity en_ZA
dc.subject Spheroids en_ZA
dc.subject Hanging drops en_ZA
dc.subject Cell lines en_ZA
dc.subject Models en_ZA
dc.subject Tissue en_ZA
dc.subject Hepatocytes en_ZA
dc.subject Liver en_ZA
dc.subject Toxicity en_ZA
dc.subject Culture en_ZA
dc.subject Tumor spheroids en_ZA
dc.subject Drug metabolism en_ZA
dc.title Characterization and reproducibility of HepG2 hanging drop spheroids toxicology in vitro en_ZA
dc.type Postprint Article en_ZA

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