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

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    Authors
    Kyffin, Jonathan A.
    Cox, Christopher R.
    Leedale, Joseph
    Colley, Helen E.
    Murdoch, Craig
    Mistry, Pratibha
    Webb, Steven D.
    Sharma, Parveen
    Publication Date
    2019-09
    
    Metadata
    Show full item record
    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. [Abstract copyright: © 2019 The Authors.]
    Citation
    Current protocols in toxicology, volume 81, issue 1, page e87
    URI
    http://hdl.handle.net/10034/622663
    DOI
    10.1002/cptx.87
    Type
    article
    Description
    From PubMed via Jisc Publications Router
    Publication status: ppublish
    Funder: BBSRC; Grant(s): BB/M503435/1
    Funder: MRC Skills Development Fellowship; Grant(s): MR/S019332/1
    Funder: University of Liverpool
    Funder: MRC
    Funder: Liverpool Centre for Mathematics in Healthcare
    Funder: EPSRC; Grant(s): EP/N014499/1
    ae974a485f413a2113503eed53cd6c53
    10.1002/cptx.87
    Scopus Count
    Collections
    Biological Sciences

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