Cytotoxicity evaluation using cryopreserved primary human hepatocytes in various culture formats


Richert, Lysiane, Baze, Audrey, Parmentier, Celine, Gerets, Helga HJ, Sison-Young, Rowena, Dorau, Martina, Lovatt, Cerys, Czich, Andreas, Goldring, Christopher, Park, B Kevin ORCID: 0000-0001-8384-824X et al (show 3 more authors) (2016) Cytotoxicity evaluation using cryopreserved primary human hepatocytes in various culture formats. TOXICOLOGY LETTERS, 258. pp. 207-215.

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Abstract

Sixteen training compounds selected in the IMI MIP-DILI consortium, 12 drug-induced liver injury (DILI) positive compounds and 4 non-DILI compounds, were assessed in cryopreserved primary human hepatocytes. When a ten-fold safety margin threshold was applied, the non-DILI-compounds were correctly identified 2h following a single exposure to pooled human hepatocytes (n=13 donors) in suspension and 14-days following repeat dose exposure (3 treatments) to an established 3D-microtissue co-culture (3D-MT co-culture, n=1 donor) consisting of human hepatocytes co-cultured with non-parenchymal cells (NPC). In contrast, only 5/12 DILI-compounds were correctly identified 2h following a single exposure to pooled human hepatocytes in suspension. Exposure of the 2D-sandwich culture human hepatocyte monocultures (2D-sw) for 3days resulted in the correct identification of 11/12 DILI-positive compounds, whereas exposure of the human 3D-MT co-cultures for 14days resulted in identification of 9/12 DILI-compounds; in addition to ximelagatran (also not identified by 2D-sw monocultures, Sison-Young et al., 2016), the 3D-MT co-cultures failed to detect amiodarone and bosentan. The sensitivity of the 2D human hepatocytes co-cultured with NPC to ximelagatran was increased in the presence of lipopolysaccharide (LPS), but only at high concentrations, therefore preventing its classification as a DILI positive compound. In conclusion (1) despite suspension human hepatocytes having the greatest metabolic capacity in the short term, they are the least predictive of clinical DILI across the MIP-DILI test compounds, (2) longer exposure periods than 72h of human hepatocytes do not allow to increase DILI-prediction rate, (3) co-cultures of human hepatocytes with NPC, in the presence of LPS during the 72h exposure period allow the assessment of innate immune system involvement of a given drug.

Item Type: Article
Uncontrolled Keywords: Cryopreserved human hepatocytes, Monoculture and co-culture, DILI, In vitro models, Short term and long term exposure
Depositing User: Symplectic Admin
Date Deposited: 03 Nov 2016 15:25
Last Modified: 19 Jan 2023 07:26
DOI: 10.1016/j.toxlet.2016.06.1127
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3004331
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