Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Sep 28, 2016; 22(36): 8178-8186
Published online Sep 28, 2016. doi: 10.3748/wjg.v22.i36.8178
Hepatocyte isolation from resected benign tissues: Results of a 5-year experience
Fan-Ying Meng, Li Liu, Jun Liu, Chun-You Li, Jian-Ping Wang, Feng-Hui Yang, Zhi-Shui Chen, Ping Zhou
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 Liu, Zhi-Shui Chen, Ping Zhou, Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Author contributions: Meng FY drafted the manuscript; Meng FY, Liu L, Li CY and Wang JP were in charge of the statistical analysis; Meng FY, Liu L, Liu J, Li CY, Wang JP and Yang FH were responsible for acquisition and analysis of the data; all authors revised and commented on the draft; all authors read and approved the final version of the manuscript.
Supported by the Major Scientific and Technological Project of Shandong Province, China, No. 201221019 and No. 2014GSF118178; the Cisco Clinical Oncology Research Fund and Bayer Schering Cancer Research Fund, No. Y-B2012-011.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board of Shandong Provincial Hospital affiliated to Shandong University.
Conflict-of-interest statement: To the best of our knowledge, no conflict of interest exists.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Feng-Hui Yang, MD, PhD, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital affiliated to Shandong University, No. 324 Jingwuweiqi Road, Jinan 250021, Shandong Province, China. yangfenghui1966@163.com
Telephone: +86-151-68889952 Fax: +86-531-68776931
Received: July 21, 2016
Peer-review started: July 22, 2016
First decision: August 29, 2016
Revised: September 1, 2016
Accepted: September 8, 2016
Article in press: September 8, 2016
Published online: September 28, 2016
Processing time: 66 Days and 15.1 Hours
Abstract
AIM

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 × 106 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 × 106 cells/g vs 3.49 ± 2.31 × 106 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 × 106 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 × 106 cells/g vs 5.31 ± 1.87 × 106 cells/g, P < 0.05).

CONCLUSION

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.

Keywords: Human hepatocyte; Primary hepatocyte; Cell isolation; Benign liver disease; Hepatocyte isolation

Core tip: We retrospectively analyzed a 5-year experience of human hepatocyte isolation from surgically-resected normal tissues and established an efficient technique for a special kind of liver samples for large-scale human 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.