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

Kyffin, JA, Cox, CR, Leedale, J, Colley, HE, Murdoch, C, Mistry, P, Webb, SD and Sharma, P (2019) Preparation of Primary Rat Hepatocyte Spheroids Utilizing the Liquid-Overlay Technique. Current Protocols in Toxicology, 81 (1). ISSN 1934-9262

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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
Uncontrolled Keywords: bile canaliculi; immunofluorescence; liquid-overlay technique; liver spheroids; primary rat hepatocytes
Subjects: R Medicine > RM Therapeutics. Pharmacology
Divisions: Applied Mathematics (merged with Comp Sci 10 Aug 20)
Publisher: Wiley
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Date Deposited: 27 Sep 2019 10:50
Last Modified: 04 Sep 2021 08:50
DOI or ID number: 10.1002/cptx.87
URI: https://researchonline.ljmu.ac.uk/id/eprint/11393
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