title: Large-scale isolation of human hepatocytes for therapeutic application.
abstract: 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 x 10(9) cells per organ, the cell preparations displaying a mean viability of 64%. The mean specific yield was 3.6 x 10(6) total and 2.6 x 10(6) 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.
authors.affiliation: Cytonet GmbH & Co KG, Branch Hannover, Hannover, Germany.
authors.forename: Krassimira
authors.initials: K
authors.lastname: Alexandrova
authors.forename: Carsten
authors.initials: C
authors.lastname: Griesel
authors.forename: Marc
authors.initials: M
authors.lastname: Barthold
authors.forename: Hans-Gert
authors.initials: HG
authors.lastname: Heuft
authors.forename: Michael
authors.initials: M
authors.lastname: Ott
authors.forename: Michael
authors.initials: M
authors.lastname: Winkler
authors.forename: Harald
authors.initials: H
authors.lastname: Schrem
authors.forename: Michael P
authors.initials: MP
authors.lastname: Manns
authors.forename: Timm
authors.initials: T
authors.lastname: Bredehorn
authors.forename: Marc
authors.initials: M
authors.lastname: Net
authors.forename: Martí Manyalich I
authors.initials: MM
authors.lastname: Vidal
authors.forename: Sabine
authors.initials: S
authors.lastname: Kafert-Kasting
authors.forename: Lubomir
authors.initials: L
authors.lastname: Arseniev
mesh_terms: D000328:Adult; D000368:Aged; D002452:Cell Count; D002469:Cell Separation / Q000379:methods* / Q000592:standards; D002470:Cell Survival; D017690:Cell Transplantation / Q000379:methods* / Q000592:standards; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods; D005234:Fatty Liver / Q000601:surgery; D005260:Female; D017408:Guidelines as Topic; D022781:Hepatocytes / Q000166:cytology* / Q000637:transplantation*; D006801:Humans; D008297:Male; D008875:Middle Aged; D011786:Quality Control; D014019:Tissue Donors; D009927:Tissue and Organ Procurement / Q000379:methods