Influence of preservation solution on the isolation and culture of human hepatocytes from liver grafts.
A major problem for the isolation and transplantation of hepatocytes is the lack of resources for obtaining viable hepatocytes. Improving this situation would enhance hepatic cell transplantation programs. Our objective was to evaluate the influence of the preservation solutions used during organ retrieval on the quality of hepatocytes isolated from liver tissue. We compared the results of the collagenase perfusion technique for isolation of hepatocytes in human livers flushed with University of Wisconsin (UW) and Celsior preservation solutions. Yield (number of viable cells per gram of tissue), cellular viability, efficiency of cells to attach to culture plates and form a monolayer, and drug metabolizing competence of the hepatocytes were measured. Successful isolation was achieved in 63% of the procedures using the UW solution and 100% of the procedures using the Celsior solution. In the UW group, significantly lower cell viability (38 +/- 41% vs. 79 +/- 14%, p < 0.05), yield of cells (4.0 +/- 5.2 x 10(6) vs. 8.2 +/- 5.6 x 10(6) cells/g, p < 0.05), and protein content at 24 h of culture (0.6 +/- 0.6 vs. 1.2 +/- 0.3 mg protein per plate, p < 0.05) than in Celsior solution were found. However, similar values of P450 activities were found in both groups. The more successful isolation, better yield, and higher cell viability obtained from human liver grafts preserved in Celsior solution, in comparison to UW solution, suggest Celsior solution as the most appropriate for preserving cadaveric hepatic tissue to be used for hepatocyte harvesting.
Serralta
Alfonso
A
Department of Surgery, Division of Liver Surgery and Transplantation, Regenerative Medicine & Cell Transplantation Unit, Hospital Universitario LA FE, Valencia, Spain. serralta_alf@gva.es
Donato
Maria Teresa
MT
Martinez
Amparo
A
Pareja
Eugenia
E
Orbis
Francisco
F
Castell
Jose Vicente
JV
Mir
Jose
J
Gómez-Lechón
Maria Jose
MJ
D000328:Adult; D000368:Aged; D002102:Cadaver; D002452:Cell Count; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D017690:Cell Transplantation; D002478:Cells, Cultured; D017364:Collagenases; D004187:Disaccharides / Q000494:pharmacology; D004573:Electrolytes / Q000494:pharmacology; D005260:Female; D005971:Glutamates / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D006639:Histidine / Q000494:pharmacology; D006801:Humans; D016031:Liver Transplantation; D008297:Male; D008353:Mannitol / Q000494:pharmacology; D008875:Middle Aged; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D013997:Time Factors
C088453:Celsior; D004187:Disaccharides; D004573:Electrolytes; D005971:Glutamates; D019188:Organ Preservation Solutions; D008353:Mannitol; D006639:Histidine; D017364:Collagenases; D005978:Glutathione