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Maximising the use of freshly isolated human hepatocytes

Evans, Peter J. 2016. Maximising the use of freshly isolated human hepatocytes. Journal of Pharmacological and Toxicological Methods 78 , pp. 85-92. 10.1016/j.vascn.2015.11.006

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Introduction Freshly isolated human hepatocytes are the best model for predicting adverse drug reactions. However, their preparation and use present the investigator with many variables that are beyond their control. These include operation continuity and timing, size and number of cut surfaces on liver tissue and the prior history of the patient. To exploit the potential of freshly isolated human hepatocytes a method is required to preserve the cells in their initial in vivo like state. This experimental pausing allows experiments to be prioritised at convenient times of the day. Methods A novel approach for selecting viable human hepatocytes by functional attachment to a gelatin gel is described rather than relying on their physical characteristics. The cells are preserved as a monolayer on the semi-solid support at 10 °C as single spherical entities. Results The hepatocytes can be released into suspension, when required, by a temperature transition to 37 °C for 20 min. The cells can be used in suspension or as a monolayer. The length of preservation depends upon the source tissue. Hepatocytes from normal liver can be maintained for at least 4 days and demonstrated to have the same level of CYP3A4 and the enzymes involved in glucuronidation and sulphation as freshly isolated cells. Cells from fatty liver, attached to gelatin, vary in their preservation time but it is at least 24 h and so confluent monolayers, that survive at 37 °C can be generated the following day. Discussion The technique enables freshly isolated human hepatocytes to be used more effectively. They can be preserved in times of plenty so more experimentation is possible. Alternatively, with poorer fatty cells the initial attachment on gelatin enables confluent monolayers of lipid rich cells to be studied.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Subjects: Q Science > QR Microbiology
Uncontrolled Keywords: Freshly isolated human hepatocytes; Preservation method; Gelatin gel
Publisher: Elsevier
ISSN: 1056-8719
Date of Acceptance: 28 November 2015
Last Modified: 28 Jun 2019 02:01

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