Kyffin, J.A., Cox, C.R., Leedale, J. et al. (5 more authors) (2019) Preparation of primary rat hepatocyte spheroids utilizing the liquid‐overlay technique. Current Protocols in Toxicology, 81 (1). e87. ISSN 1934-9254
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.
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
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2019 The Authors. This is an open access article under the terms of the Creative Commons Attribution License, http://creativecommons.org/licenses/by/4.0/, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
Keywords: | bile canaliculi; immunofluorescence; liquid‐overlay technique; liver spheroids; primary rat hepatocytes |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > School of Clinical Dentistry (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 28 Nov 2019 14:19 |
Last Modified: | 28 Nov 2019 14:19 |
Status: | Published |
Publisher: | Wiley |
Refereed: | Yes |
Identification Number: | 10.1002/cptx.87 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:153760 |
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Filename: Kyffin_et_al-2019-Current_Protocols_in_Toxicology.pdf
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