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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.

Lee SM, Schelcher C, Laubender RP, Fröse N, Thasler RM, Schiergens TS, Mansmann U, Thasler WE - PLoS ONE (2014)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of General, Visceral, Transplantation, Vascular and Thoracic Surgery, Grosshadern Hospital, Ludwig Maximilians University, Munich, Germany.

ABSTRACT
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.

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Related in: MedlinePlus

Variables measured in the blood or serum that have significant relationships with the viability (%) of hepatocytes after linear regression analyses.Figures show relationships between viability and (A) aspartate aminotransferase activity (GOT; U/L), (B) gamma-glutamyltranspeptidase activity (GGT; U/L), (C) alanine aminotransferase activity (GPT; U/L), (D) bilirubin (mg/dL) or (E) quick value (%). Values were deemed significant when P<0.05.
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pone-0107567-g002: Variables measured in the blood or serum that have significant relationships with the viability (%) of hepatocytes after linear regression analyses.Figures show relationships between viability and (A) aspartate aminotransferase activity (GOT; U/L), (B) gamma-glutamyltranspeptidase activity (GGT; U/L), (C) alanine aminotransferase activity (GPT; U/L), (D) bilirubin (mg/dL) or (E) quick value (%). Values were deemed significant when P<0.05.


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

Lee SM, Schelcher C, Laubender RP, Fröse N, Thasler RM, Schiergens TS, Mansmann U, Thasler WE - PLoS ONE (2014)

Variables measured in the blood or serum that have significant relationships with the viability (%) of hepatocytes after linear regression analyses.Figures show relationships between viability and (A) aspartate aminotransferase activity (GOT; U/L), (B) gamma-glutamyltranspeptidase activity (GGT; U/L), (C) alanine aminotransferase activity (GPT; U/L), (D) bilirubin (mg/dL) or (E) quick value (%). Values were deemed significant when P<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196847&req=5

pone-0107567-g002: Variables measured in the blood or serum that have significant relationships with the viability (%) of hepatocytes after linear regression analyses.Figures show relationships between viability and (A) aspartate aminotransferase activity (GOT; U/L), (B) gamma-glutamyltranspeptidase activity (GGT; U/L), (C) alanine aminotransferase activity (GPT; U/L), (D) bilirubin (mg/dL) or (E) quick value (%). Values were deemed significant when P<0.05.
Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of General, Visceral, Transplantation, Vascular and Thoracic Surgery, Grosshadern Hospital, Ludwig Maximilians University, Munich, Germany.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus