Characterisation of a functional rat hepatocyte spheroid model

Kyffin, Jonathan A, Sharma, Parveen ORCID: 0000-0002-5534-2417, Leedale, Joseph ORCID: 0000-0001-9010-4126, Colley, Helen E, Murdoch, Craig, Harding, Amy L, Mistry, Pratibha and Webb, Steven D
(2019) Characterisation of a functional rat hepatocyte spheroid model. TOXICOLOGY IN VITRO, 55. pp. 160-172.

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Many in vitro liver cell models, such as 2D systems, that are used to assess the hepatotoxic potential of xenobiotics suffer major limitations arising from a lack of preservation of physiological phenotype and metabolic competence. To circumvent some of these limitations there has been increased focus on producing more representative 3D models. Here we have used a novel approach to construct a size-controllable 3D hepatic spheroid model using freshly isolated primary rat hepatocytes (PRH) utilising the liquid-overlay technique whereby PRH spontaneously self-assemble in to 3D microtissues. This system produces viable spheroids with a compact in vivo-like structure for up to 21 days with sustained albumin production for the duration of the culture period. F-actin was seen throughout the spheroid body and P-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (MRP2) transporters had polarised expression on the canalicular membrane of hepatocytes within the spheroids upon formation (day 3). The MRP2 transporter was able to functionally transport 5 μM 5-chloromethylfluorescein diacetate (CMFDA) substrates into these canalicular structures. These PRH spheroids display in vivo characteristics including direct cell-cell contacts, cellular polarisation, 3D cellular morphology, and formation of functional secondary structures throughout the spheroid. Such a well-characterised system could be readily exploited for pre-clinical and non-clinical repeat-dose investigations and could make a significant contribution to replace, reduce and refine the use of animals for applied research.

Item Type: Article
Uncontrolled Keywords: Liver spheroids, 3D cell culture, Primary rat hepatocytes, Polarisation, Bile canaliculi, Liquid-overlay technique
Depositing User: Symplectic Admin
Date Deposited: 07 Feb 2019 08:30
Last Modified: 19 Jan 2023 01:08
DOI: 10.1016/j.tiv.2018.12.014
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