Hepatocyte isolation from resected benign tissues: Results of a 5-year experience.
OBJECTIVE
To analyze retrospectively a 5-year experience of human hepatocyte isolation from resected liver tissues with benign disease.


METHODS
We established a method of modified four-step retrograde perfusion to isolate primary human hepatocytes. Samples were collected from the resected livers of patients with intrahepatic duct calculi (n = 7) and liver hemangioma (n = 17). Only the samples weighing ≥ 15 g were considered suitable for hepatocyte isolation. By using the standard trypan blue exclusion technique, hepatocyte viability and yield were immediately determined after isolation.


RESULTS
Twenty-four liver specimens, weighing 15-42 g, were immediately taken from the margin of the removed samples and transferred to the laboratory for hepatocyte isolation. Warm ischemia time was 5-35 min and cold ischemia time was 15-45 min. For the 7 samples of intrahepatic duct calculi, the method resulted in a hepatocyte yield of 3.49 ± 2.31 × 10(6) hepatocytes/g liver, with 76.4% ± 10.7% viability. The 17 samples of liver hemangioma had significantly higher yield of cells (5.4 ± 1.71 × 10(6) cells/g vs 3.49 ± 2.31 × 10(6) cells/g, P < 0.05) than the samples of intrahepatic duct calculi. However, there seems to be no clear difference in cell viability (80.3% ± 9.67% vs 76.4% ± 10.7%, P > 0.05). We obtained a cell yield of 5.31 ± 1.87 × 10(6) hepatocytes/g liver when the samples weighed > 20 g. However, for the tissues weighing ≤ 20 g, a reduction in yield was found (3.08 ± 1.86 × 10(6) cells/g vs 5.31 ± 1.87 × 10(6) cells/g, P < 0.05).


CONCLUSIONS
Benign diseased livers are valuable sources for large-number hepatocyte isolation. Our study represents the largest number of primary human hepatocytes isolated from resected specimens from patients with benign liver disease. We evaluated the effect of donor liver characteristics on cell isolation, and we found that samples of liver hemangioma can provide better results than intrahepatic duct calculi, in terms of cell yield. Furthermore, the size of the tissues can affect the outcome of hepatocyte isolation.
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Fan-Ying
FY
Meng
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Li
L
Liu
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Jun
J
Liu
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Chun-You
CY
Li
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Jian-Ping
JP
Wang
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Feng-Hui
FH
Yang
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Zhi-Shui
ZS
Chen
Fan-Ying Meng, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Ping
P
Zhou
D000328:Adult; D000368:Aged; D002137:Calculi / Q000601:surgery; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D005260:Female; D006391:Hemangioma / Q000601:surgery; D006500:Hepatic Duct, Common / Q000601:surgery; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000503:physiopathology / Q000601:surgery*; D008297:Male; D008875:Middle Aged; D010477:Perfusion; D012189:Retrospective Studies; D052096:Warm Ischemia
Benign liver disease; Cell isolation; Hepatocyte isolation; Human hepatocyte; Primary hepatocyte
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