Preparation of Primary Rat Hepatocyte Spheroids Utilizing the Liquid-Overlay Technique.



Kyffin, Jonathan A, Cox, Christopher R, Leedale, Joseph ORCID: 0000-0001-9010-4126, Colley, Helen E, Murdoch, Craig, Mistry, Pratibha, Webb, Steven D and Sharma, Parveen ORCID: 0000-0002-5534-2417
(2019) Preparation of Primary Rat Hepatocyte Spheroids Utilizing the Liquid-Overlay Technique. Current protocols in toxicology, 81 (1). e87 - e87.

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Abstract

Herein, we describe a protocol for the preparation and analysis of primary isolated rat hepatocytes in a 3D cell culture format described as spheroids. The hepatocyte cells spontaneously self-aggregate into spheroids without the need for synthetic extracellular matrices or hydrogels. Primary rat hepatocytes (PRHs) are a readily available source of primary differentiated liver cells and therefore conserve many of the required liver-specific functional markers, and elicit the natural in vivo phenotype when compared with common hepatic cells lines. We describe the liquid-overlay technique which provides an ultra-low attachment surface on which PRHs can be cultured as spheroids. © 2019 The Authors. Basic Protocol 1: Preparation of agarose-coated plates Basic Protocol 2: Primary rat hepatocyte isolation procedure Basic Protocol 3: Primary rat hepatocyte spheroid culture Basic Protocol 4: Immunofluorescent analysis of PRH spheroids.

Item Type: Article
Depositing User: Symplectic Admin
Date Deposited: 19 Sep 2019 11:45
Last Modified: 09 Jan 2021 03:09
DOI: 10.1002/cptx.87
URI: https://livrepository.liverpool.ac.uk/id/eprint/3055199