dc.contributor.author |
Hurrell, Tracey
|
|
dc.contributor.author |
Ellero, Andrea Antonio
|
|
dc.contributor.author |
Masso, Zelie Flavienne
|
|
dc.contributor.author |
Cromarty, Allan Duncan
|
|
dc.date.accessioned |
2018-08-07T10:02:06Z |
|
dc.date.issued |
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 |
http://www.elsevier.com/locate/tiv |
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.identifier.uri |
http://hdl.handle.net/2263/66119 |
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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 |