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Large-Scale Isolation of Human Hepatocytes for Therapeutic Application

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During the last decade, hepatocyte transplantation has been suggested as a safe and potentially effective clinical option for the treatment of acute or decompensating chronic liver failure as well as for hereditary liver disease. Currently, one of the major limiting factors for clinical application is the insufficient access to suitable liver cell preparations. In cooperation with the German and Catalane organ procurement organizations, a routine procedure for the isolation of hepatocytes from donor organs rejected for transplantation (n = 117) has been established. The process is performed according to the current EC Guidelines for Good Manufacturing Practice (cGMP) and all corresponding national laws and regulations concerning donor organ and tissue procurement. In about 50% of the cases (n = 58) the three-step perfusion procedure has been completed with an average total cell yield of 5.9 × 109 cells per organ, the cell preparations displaying a mean viability of 64%. The mean specific yield was 3.6 × 106 total and 2.6 × 106 viable cells per gram liver tissue, respectively. Specific cell yields from three infantile donor livers were considerably higher. No correlation between isolation efficiency and cold ischemia time or donor age was found within the adult organ donors. In contrast, organs with a severe steatosis generally did not result in successful cell isolation. Results of sterility and endotoxin determination are also presented. In summary, a standardized and cGMP conform method of hepatocyte isolation from nontransplantable liver organs was established, which reproducibly yields large amounts of hepatocytes suitable for therapeutic application.
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Keywords: Cell transplantation; GMP; Hepatocyte isolation; Steatosis

Document Type: Research Article

Affiliations: 1: *Cytonet GmbH & Co. KG, Branch Hannover, Feodor-Lynen-Str. 21, D-30625 Hannover, Germany 2: Cytonet GmbH & Co. KG, Branch Hannover, Feodor-Lynen-Str. 21, D-30625 Hannover, Germany 3: Cytonet GmbH & Co. KG, Branch Hannover, Feodor-Lynen-Str. 21, D-30625 Hannover, Germany, Department of Transfusion Medicine, Hannover Medical School, Carl-Neuberg-Str. 1, D-30625 Hannover, Germany 4: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Carl-Neuberg-Str. 1, D-30625 Hannover, Germany 5: Department of Visceral and Transplantation Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, D-30625 Hannover, Germany 6: DSO-G, Feodor-Lynen-Str. 21, D-30625 Hannover, Germany 7: Fundació Clínic per a la Recerca Biomédica, Hospital Clínic Barcelona, Villarroel, 170, E-08036 Barcelona, Spain 8: Cytonet GmbH & Co. KG, Branch Hannover, Feodor-Lynen-Str. 21, D-30625 Hannover, Germany, Department of Hematology and Oncology, Hannover Medical School, Carl-Neuberg-Str. 1, D-30625 Hannover, Germany

Publication date: 2005-10-01

More about this publication?
  • Cell Transplantation publishes original, peer-reviewed research and review articles on the subject of cell transplantation and its application to human diseases. To ensure high-quality contributions from all areas of transplantation, separate section editors and editorial boards have been established. Articles deal with a wide range of topics including physiological, medical, preclinical, tissue engineering, and device-oriented aspects of transplantation of nervous system, endocrine, growth factor-secreting, bone marrow, epithelial, endothelial, and genetically engineered cells, among others. Basic clinical studies and immunological research papers are also featured. To provide complete coverage of this revolutionary field, Cell Transplantation will report on relevant technological advances, and ethical and regulatory considerations of cell transplants. Cell Transplantation is now an Open Access journal starting with volume 18 in 2009, and therefore there will be an inexpensive publication charge, which is dependent on the number of pages, in addition to the charge for color figures. This will allow work to be disseminated to a wider audience and also entitle the corresponding author to a free PDF, as well as prepublication of an unedited version of the manuscript.

    Cell Transplantation is now being published by SAGE. Please visit their website for the most recent issues.

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