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 |
|---|---|
| Uncontrolled Keywords: | Spheroids, Cellular, Hepatocytes, Animals, Rats, Culture Media, Cell Culture Techniques |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 19 Sep 2019 11:45 |
| Last Modified: | 19 Jan 2023 00:26 |
| DOI: | 10.1002/cptx.87 |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3055199 |
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