Cytotoxicity evaluation using cryopreserved primary human hepatocytes in various culture formats

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• 作者：Lysiane Richert^a ; ^h ; ^{l.richert@kaly-cell.com" class="auth_mail" title="E-mail the corresponding author} ; Audrey Baze^a ; ^{a.baze@kaly-cell.com" class="auth_mail" title="E-mail the corresponding author} ; Cé ; line Parmentier^a ; ^{c.parmentier@kaly-cell.com" class="auth_mail" title="E-mail the corresponding author} ; Helga H.J. Gerets^b ; ^{helga.gerets@ucb.com" class="auth_mail" title="E-mail the corresponding author} ; Rowena Sison-Young^c ; ^{Rowena.Sison-Young@liverpool.ac.uk" class="auth_mail" title="E-mail the corresponding author} ; Martina Dorau^d ; ^{Martina.Dorau@sanofi.com" class="auth_mail" title="E-mail the corresponding author} ; Cerys Lovatt^e ; ^{cerys.a.lovatt@gsk.com" class="auth_mail" title="E-mail the corresponding author} ; Andreas Czich^d ; ^{Andreas.Czich@sanofi.com" class="auth_mail" title="E-mail the corresponding author} ; Christopher Goldring^c ; ^{C.E.P.Goldring@liverpool.ac.uk" class="auth_mail" title="E-mail the corresponding author} ; B. Kevin Park^c ; ^{B.K.Park@liv.ac.uk" class="auth_mail" title="E-mail the corresponding author} ; Satu Juhila^f ; ^{satu.juhila@orionpharma.com" class="auth_mail" title="E-mail the corresponding author} ; Alison J. Foster^g ; ^{Alison.Foster2@astrazeneca.com" class="auth_mail" title="E-mail the corresponding author} ; Dominic P. Williams^g ; ^{Dominic.P.Williams@astrazeneca.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：DILI ; drug-induced liver injury ; NPC ; non-parenchymal cells ; 2D-sw ; 2D-sandwich ; SM ; safety margin ; MT ; microtissue ; TC ; training compound
• 刊名：Toxicology Letters
• 出版年：2016
• 出版时间：6 September 2016
• 年：2016
• 卷：258
• 期：Complete
• 页码：207-215
• 全文大小：2392 K

文摘

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 2 h 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 2 h following a single exposure to pooled human hepatocytes in suspension. Exposure of the 2D-sandwich culture human hepatocyte monocultures (2D-sw) for 3 days resulted in the correct identification of 11/12 DILI-positive compounds, whereas exposure of the human 3D-MT co-cultures for 14 days 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 72 h 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 72 h exposure period allow the assessment of innate immune system involvement of a given drug.