Preparation of Primary Rat Hepatocyte Spheroids Utilizing the Liquid-Overlay Technique
Authors
Kyffin, Jonathan A.Cox, Christopher R.
Leedale, Joseph
Colley, Helen E.
Murdoch, Craig
Mistry, Pratibha
Webb, Steven D.
Sharma, Parveen
Affiliation
University of Liverpool; University of Chester; University of Sheffield; Syngenta Ltd; Liverpool John Moores UniversityPublication Date
2019-09-13
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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. [Abstract copyright: © 2019 The Authors.]Citation
Kyffin, J. A., Cox, C. R., Leedale, J., Colley, H. E., Murdoch, C., Mistry, P., Webb, S. D., & Sharma, P. (2019). Preparation of primary rat hepatocyte spheroids utilizing the liquid-overlay technique. Current Protocols in Toxicology, 81(1), e87. https://doi.org/10.1002/cptx.87Publisher
Wiley-BlackwellJournal
Current Protocols in ToxicologyDOI
10.1002/cptx.87Additional Links
https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cptx.87Type
ArticleISSN
1934-9254EISSN
1934-9262ae974a485f413a2113503eed53cd6c53
10.1002/cptx.87
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Except where otherwise noted, this item's license is described as https://creativecommons.org/licenses/by/4.0/