Hepatocyte isolation from resected benign tissues: Results of a 5-year experience

High levels of reactive oxygen species (ROS) can lead to impairment of cell structure, biomolecules’ loss of function and cell death and are associated with liver diseases. Cells that survive increased ROS often undergo malignant transformation. Many cancer cells tolerate high levels of ROS. Here we report a transiently increased production of H2O2 and concomitant upregulation of antioxidative enzymes triggered by hepatocyte isolation; the H2O2 levels revert in about two days in culture. Three-day survival rate of the isolated cells in the presence of 2.5-fold increase of H2O2 is almost 80%. Apoptosis activation through the mitochondrial pathway is meanwhile reduced by inhibition of caspase-9 triggering. This reduction depends on the amount of H2O2 production, as decreased production of H2O2 in the presence of an antioxidant results in increased apoptosis triggering. These stress adaptations do not influence urea production, which is unchanged throughout the normal and stress adapted phases. We conclude that hepatocytes’ stress adaptation is mediated by increased ROS production. In this case, high ROS improve cell survival.

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Nonenzymatic versus Enzymatic Hepatocyte Isolation from Pig Livers for Larger Scale Investigations of Liver Cell Perfusion Systems

The International Journal of Artificial Organs ◽

10.1177/039139889301600909 ◽

1993 ◽

Vol 16 (9) ◽

pp. 677-681 ◽

Cited By ~ 20

Author(s):

J.C. Gerlach ◽

J. Brombacher ◽

J.M. Courtney ◽

P. Neuhaus

Keyword(s):

Liver Cell ◽

Perfusion Technique ◽

Hepatocyte Cultures ◽

Hepatocyte Isolation ◽

Collagenase Perfusion

A comparison of nonenzymatic and enzymatic hepatocyte isolation was performed on pig livers. The collagenase perfusion showed superior results: mean viability 72 ± 10% versus a maximum viability of 21% using EDTA-perfusion. A five-step collagenase perfusion technique, developed for pig livers enables larger scale investigations, in order to develop methods for hepatocyte cultures in therapeutical liver cell perfusion systems.

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Predictive value of the human hepatocyte isolation results for postoperative morbidity after major hepatectomy

HPB ◽

10.1016/j.hpb.2016.01.065 ◽

2016 ◽

Vol 18 ◽

pp. e686

Author(s):

F. Le Roux ◽

Y.E. Herpe ◽

A. Bengrine ◽

A.S. Bruyer ◽

J.M. Regimbeau

Keyword(s):

Predictive Value ◽

Postoperative Morbidity ◽

Major Hepatectomy ◽

Human Hepatocyte ◽

Hepatocyte Isolation

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Experience of Microbiological Screening of Human Hepatocytes for Clinical Transplantation

Cell Transplantation ◽

10.3727/096368909x471323 ◽

2009 ◽

Vol 18 (8) ◽

pp. 941-947 ◽

Cited By ~ 13

Author(s):

Sharon C. Lehec ◽

Robin D. Hughes ◽

Ragai R. Mitry ◽

Michelle A. Graver ◽

Anita Verma ◽

...

Keyword(s):

Liver Tissue ◽

Metabolic Disorders ◽

Bacterial Contamination ◽

Coagulase Negative Staphylococci ◽

Culture Bottle ◽

Clinical Transplantation ◽

Hepatocyte Isolation ◽

Human Use ◽

Initial Contamination ◽

Preservative Solutions

Hepatocyte transplantation is being used in patients with liver-based metabolic disorders and acute liver failure. Hepatocytes are isolated from unused donor liver tissue under GMP conditions. Cells must be free of microbiological contamination to be safe for human use. The experience of microbiological screening during 72 hepatocyte isolation procedures at one center is reported. Samples were taken at different stages of the process and tested using a blood culture bottle system and Gram stain. Bacterial contamination was detected in 37.5% of the UW organ preservative solutions used to transport the liver tissue to the Cell Isolation Unit. After tissue processing the contamination was reduced to 7% overall in the final hepatocyte product, irrespective of the presence of initial contamination of the transport solution. The most common organisms recovered were coagulase-negative staphylococci, a skin commensal. A total of 41 preparations of fresh or cryopreserved hepatocytes were used for cell transplantation in children with liver-based metabolic disorders without any evidence of sepsis due to infusion of hepatocytes. In conclusion, the incidence of bacterial contamination of the final product was low, confirming the suitability of the organs used, hepatocyte isolation procedure, and the environmental conditions of the clean room.

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