title: Characterization of a functional C3A liver spheroid model.
abstract: More predictivein vitroliver models are a critical requirement for preclinical screening of compounds demonstrating hepatotoxic liability. 3D liver spheroids have been shown to have an enhanced functional lifespan compared to 2D monocultures; however a detailed characterisation of spatiotemporal function and structure of spheroids still needs further attention before widespread use in industry. We have developed and characterized the structure and function of a 3D liver spheroid model formed from C3A hepatoma cells. Spheroids were viable and maintained a compactin vivo-like structure with zonation features for up to 32 days. MRP2 and Pgp transporters had polarised expression on the canalicular membrane of cells in the spheroids and were able to functionally transport CMFDA substrate into these canalicular structures. Spheroids expressed CYP2E1 and were able to synthesise and secrete albumin and urea to a higher degree than monolayer C3A cultures. Penetration of doxorubicin throughout the spheroid core was demonstrated. Spheroids showed increased susceptibility to hepatotoxins when compared to 2D cultures, with acetaminophen having an IC50of 7.2 mM in spheroids compared to 33.8 mM in monolayer culture. To conclude, we developed an alternative method for creating C3A liver spheroids and demonstrated cellular polarisation and zonation, as well as superior liver-specific functionality and more sensitive toxicological response compared to standard 2D liver models, confirming a morein vivo-like liver model.
authors.affiliation: MRC Centre for Drug Safety Science , Department of Molecular and Clinical Pharmacology , Sherrington Building , Ashton Street and University of Liverpool , L69 3GE , UK . Email: parveen.sharma@liverpool.ac.uk; AstraZeneca , 310 , Cambridge Science Park , Milton Road , Cambridge , Cambridgeshire , CB4 0FZ , UK.
authors.forename: Harriet
authors.initials: H
authors.lastname: Gaskell
authors.affiliation: MRC Centre for Drug Safety Science , Department of Molecular and Clinical Pharmacology , Sherrington Building , Ashton Street and University of Liverpool , L69 3GE , UK . Email: parveen.sharma@liverpool.ac.uk.
authors.forename: Parveen
authors.initials: P
authors.lastname: Sharma
authors.affiliation: Academic Unit of Oral and Maxillofacial Pathology , School of Clinical Dentistry , Claremont Crescent and University of Sheffield , Sheffield , S10 2TA , UK.
authors.forename: Helen E
authors.initials: HE
authors.lastname: Colley
authors.affiliation: Academic Unit of Oral and Maxillofacial Pathology , School of Clinical Dentistry , Claremont Crescent and University of Sheffield , Sheffield , S10 2TA , UK.
authors.forename: Craig
authors.initials: C
authors.lastname: Murdoch
authors.affiliation: AstraZeneca , 310 , Cambridge Science Park , Milton Road , Cambridge , Cambridgeshire , CB4 0FZ , UK.
authors.forename: Dominic P
authors.initials: DP
authors.lastname: Williams
authors.affiliation: Department of Mathematical Sciences , Liverpool John Moores University , James Parsons Building , Byrom Street , Liverpool , L3 3AF , UK.
authors.forename: Steven D
authors.initials: SD
authors.lastname: Webb