Journal article
The isolation of primary hepatocytes from human tissue: optimising the use of small non-encapsulated liver resection surplus
- Abstract:
- Two-step perfusion is considered the gold standard method for isolating hepatocytes from human liver tissue. As perfusion may require a large tissue specimen, which is encapsulated and has accessible vessels for cannulation, only a limited number of tissue samples may be suitable. Therefore, the aim of this work was to develop an alternative method to isolate hepatocytes from non-encapsulated and small samples of human liver tissue. Healthy tissue from 44 human liver resections were graded for steatosis and tissue weights between 7.8 and 600 g were used for hepatocyte isolations. Tissue was diced and underwent a two-step digestion (EDTA and collagenase). Red cell lysis buffer was used to prevent red blood cell contamination and toxicity. Isolated hepatocyte viability was determined by trypan blue exclusion. Western blot and biochemical analyses were undertaken to ascertain cellular phenotype and function. Liver tissue that weighed ≥50 g yielded significantly higher (P < 0.01) cell viability than tissue <50 g. Viable cells secreted urea and displayed the phenotypic hepatocyte markers albumin and cytochrome P450. Presence of steatosis in liver tissue or intra-hepatocellular triglyceride content had no effect on cell viability. This methodology allows for the isolation of viable primary human hepatocytes from small amounts of “healthy” resected liver tissue which are not suitable for perfusion. This work provides the opportunity to increase the utilisation of resection surplus tissue, and may ultimately lead to an increased number of in vitro cellular studies being undertaken using the gold-standard model of human primary hepatocytes.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 454.3KB, Terms of use)
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- Publisher copy:
- 10.1007/s10561-017-9641-6
Authors
+ British Heart Foundation
More from this funder
- Funding agency for:
- Hodson, L
- Grant:
- Senior Fellow in Basic Science
- FS/11/18/28633
- Publisher:
- Springer Netherlands
- Journal:
- Cell and Tissue Banking More from this journal
- Volume:
- 18
- Issue:
- 4
- Pages:
- 597–604
- Publication date:
- 2017-07-17
- Acceptance date:
- 2017-07-06
- DOI:
- EISSN:
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1573-6814
- ISSN:
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1389-9333
- Keywords:
- Pubs id:
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pubs:704612
- UUID:
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uuid:2e2ca993-dabc-4977-80f5-ff3c39e520d9
- Local pid:
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pubs:704612
- Source identifiers:
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704612
- Deposit date:
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2017-07-07
Terms of use
- Copyright holder:
- Green et al
- Copyright date:
- 2017
- Notes:
- © The Authors 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
- Licence:
- CC Attribution (CC BY)
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