An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.

Isolated human primary hepatocytes are an essential in vitro model for basic and clinical research. For successful application as a model, isolated hepatocytes need to have a good viability and be available in sufficient yield. Therefore, this study aims to identify donor characteristics, intra-oper...

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Autores principales:	Serene M L Lee, Celine Schelcher, Rüdiger P Laubender, Natalja Fröse, Reinhard M K Thasler, Tobias S Schiergens, Ulrich Mansmann, Wolfgang E Thasler
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/96e88cdca97a4b6ba9ed5ea883dbc286
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spelling	oai:doaj.org-article:96e88cdca97a4b6ba9ed5ea883dbc2862021-11-25T05:56:42ZAn algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.1932-620310.1371/journal.pone.0107567https://doaj.org/article/96e88cdca97a4b6ba9ed5ea883dbc2862014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0107567https://doaj.org/toc/1932-6203Isolated human primary hepatocytes are an essential in vitro model for basic and clinical research. For successful application as a model, isolated hepatocytes need to have a good viability and be available in sufficient yield. Therefore, this study aims to identify donor characteristics, intra-operative factors, tissue processing and cell isolation parameters that affect the viability and yield of human hepatocytes. Remnant liver pieces from tissue designated as surgical waste were collected from 1034 donors with informed consent. Human hepatocytes were isolated by a two-step collagenase perfusion technique with modifications and hepatocyte yield and viability were subsequently determined. The accompanying patient data was collected and entered into a database. Univariate analyses found that the viability and the yield of hepatocytes were affected by many of the variables examined. Multivariate analyses were then carried out to confirm the factors that have a significant relationship with the viability and the yield. It was found that the viability of hepatocytes was significantly decreased by the presence of fibrosis, liver fat and with increasing gamma-glutamyltranspeptidase activity and bilirubin content. Yield was significantly decreased by the presence of liver fat, septal fibrosis, with increasing aspartate aminotransferase activity, cold ischemia times and weight of perfused liver. However, yield was significantly increased by chemotherapy treatment. In conclusion, this study determined the variables that have a significant effect on the viability and the yield of isolated human hepatocytes. These variables have been used to generate an algorithm that can calculate projected viability and yield of isolated human hepatocytes. In this way, projected viability can be determined even before isolation of hepatocytes, so that donors that result in high viability and yield can be identified. Further, if the viability and yield of the isolated hepatocytes is lower than expected, this will highlight a methodological problem that can be addressed.Serene M L LeeCeline SchelcherRüdiger P LaubenderNatalja FröseReinhard M K ThaslerTobias S SchiergensUlrich MansmannWolfgang E ThaslerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 10, p e107567 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Serene M L Lee Celine Schelcher Rüdiger P Laubender Natalja Fröse Reinhard M K Thasler Tobias S Schiergens Ulrich Mansmann Wolfgang E Thasler An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.
description	Isolated human primary hepatocytes are an essential in vitro model for basic and clinical research. For successful application as a model, isolated hepatocytes need to have a good viability and be available in sufficient yield. Therefore, this study aims to identify donor characteristics, intra-operative factors, tissue processing and cell isolation parameters that affect the viability and yield of human hepatocytes. Remnant liver pieces from tissue designated as surgical waste were collected from 1034 donors with informed consent. Human hepatocytes were isolated by a two-step collagenase perfusion technique with modifications and hepatocyte yield and viability were subsequently determined. The accompanying patient data was collected and entered into a database. Univariate analyses found that the viability and the yield of hepatocytes were affected by many of the variables examined. Multivariate analyses were then carried out to confirm the factors that have a significant relationship with the viability and the yield. It was found that the viability of hepatocytes was significantly decreased by the presence of fibrosis, liver fat and with increasing gamma-glutamyltranspeptidase activity and bilirubin content. Yield was significantly decreased by the presence of liver fat, septal fibrosis, with increasing aspartate aminotransferase activity, cold ischemia times and weight of perfused liver. However, yield was significantly increased by chemotherapy treatment. In conclusion, this study determined the variables that have a significant effect on the viability and the yield of isolated human hepatocytes. These variables have been used to generate an algorithm that can calculate projected viability and yield of isolated human hepatocytes. In this way, projected viability can be determined even before isolation of hepatocytes, so that donors that result in high viability and yield can be identified. Further, if the viability and yield of the isolated hepatocytes is lower than expected, this will highlight a methodological problem that can be addressed.
format	article
author	Serene M L Lee Celine Schelcher Rüdiger P Laubender Natalja Fröse Reinhard M K Thasler Tobias S Schiergens Ulrich Mansmann Wolfgang E Thasler
author_facet	Serene M L Lee Celine Schelcher Rüdiger P Laubender Natalja Fröse Reinhard M K Thasler Tobias S Schiergens Ulrich Mansmann Wolfgang E Thasler
author_sort	Serene M L Lee
title	An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.
title_short	An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.
title_full	An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.
title_fullStr	An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.
title_full_unstemmed	An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.
title_sort	algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/96e88cdca97a4b6ba9ed5ea883dbc286
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An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.

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