Isolation of primary human hepatocytes after partial hepatectomy: criteria for identification of the most promising liver specimen.
Demands for primary human hepatocytes are continuously increasing, while supply is insufficient due to limited cell sources. To improve cell availability, the present study investigates the influence of donor liver characteristics on the outcome of hepatocyte isolation from surgically removed liver tissue (n = 50). Hepatocytes were isolated from liver specimens using a standardized two-step collagenase perfusion technique. The patient's sex, previous chemotherapy, or histopathology have shown no influence. Donor age significantly affected the isolation outcome, but was not found suitable for predicting cell yields. Preoperative blood parameters did not correlate with cell yield, although cell function was affected: total protein, albumin synthesis, and cell viability were significantly decreased for serum gamma-glutamyl-transferase (GGT) levels >60 U/L. Specimens from patients with benign diseases gave significantly higher cell yields than tissue removed due to secondary and primary tumors, respectively. The indication for surgery is a valuable basis for identifying the most yielding specimens. Hepatocytes from donors with high GGT levels appear to show reduced functional properties.
Vondran
Florian Wolfgang Rudolf
FW
Department of General, Visceral, and Transplantation Surgery, Charité Universitätsmedizin Berlin, Campus Virchow, Berlin, Germany. florian@vondran.org
Katenz
Ekaterina
E
Schwartlander
Ruth
R
Morgul
Mehmet Haluk
MH
Raschzok
Nathanael
N
Gong
Xiaobing
X
Cheng
Xiangdong
X
Kehr
Daniel
D
Sauer
Igor Maximilian
IM
D000328:Adult; D000368:Aged; D000418:Albumins / Q000096:biosynthesis; D001219:Aspartate Aminotransferases / Q000378:metabolism; D018929:Cell Culture Techniques; D002469:Cell Separation; D048430:Cell Shape; D002470:Cell Survival; D002478:Cells, Cultured; D005260:Female; D006498:Hepatectomy; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000201:enzymology / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D008297:Male; D008875:Middle Aged; D014176:Protein Biosynthesis; D013997:Time Factors; D014508:Urea / Q000378:metabolism; D005723:gamma-Glutamyltransferase / Q000097:blood
D000418:Albumins; D014508:Urea; D007770:L-Lactate Dehydrogenase; D005723:gamma-Glutamyltransferase; D001219:Aspartate Aminotransferases