Rapid Communication
Copyright ©2008 The WJG Press and Baishideng. All rights reserved.
World J Gastroenterol. Aug 28, 2008; 14(32): 5025-5031
Published online Aug 28, 2008. doi: 10.3748/wjg.14.5025
Hepatocytes isolated from neoplastic liver-immunomagnetic purging as a new source for transplantation
Aravin Gunasegaram, Javed Akhter, Peng Yao, Loreena A Johnson, Stephen M Riodan, David L Morris
Aravin Gunasegaram, Javed Akhter, Peng Yao, Loreena A Johnson, David L Morris, University of New South Wales, Department of Surgery, St George Hospital, Sydney 2044, Australia
Stephen M Riodan, Gastrointestinal and Liver Unit, Prince of Wales Hospital, University of New South Wales, Sydney 2044, Australia
Author contributions: Gunasegaram A, Akhter J and Johnson LA contributed equally to this work; Akhter J and Morris DL designed research; Gunasegaram A, Akhter J and Johnson LA performed research; Gunasegaram A and Akhter J analysed data; Gunasegaram A, Akhter J, Yao P, Johnson LA and Riodan SM wrote the paper.
Correspondence to: David L Morris, Professor, Level 3 Pitney Building, St George Hospital, Kogarah 2217, New South Wales, Australia. david.morris@unsw.edu.au
Telephone: +61-2-93502070 Fax: +61-2-93503997
Received: February 1, 2008
Revised: August 8, 2008
Accepted: August 15, 2008
Published online: August 28, 2008
Abstract

AIM: To investigate whether hepatocytes isolated from macroscopically normal liver during hepatic resection for neoplasia could provide a novel source of healthy hepatocytes, including the development of reliable protocols for malignant cells removal from the hepatocyte preparation.

METHODS: Hepatocytes were procured from resected liver of 18 patients with liver tumors using optimised digestion and cell-enrichment protocols. Suspensions of various known quantities of the HT-29 tumor cell line and patient hepatocytes were treated or not with Ep-CAM-antibody-coated immunomagnetic beads in order to investigate the efficacy of tumor-purging by immunomagnetic depletion, using a semi-quantitative RT-PCR method developed to detect tumor cells. Immunomagnetic bead-treated or bead-untreated tumor cell-hepatocyte suspensions were transplanted intra-peritoneally in Balb/C nude mice to assess the rates of tumor development.

RESULTS: Mean viable hepatocyte yield was 9.3 x 106 cells per gram of digested liver with mean viability of 70.5%. Immunomagnetic depletion removed tumor cells to below the RT-PCR detection-threshold of 1 tumor cell in 106 hepatocytes, representing a maximum tumor purging efficacy of greater than 400 000-fold. Transplanted, immunomagnetic bead-purged tumor cell-hepatocyte suspensions did not form peritoneal tumors in Balb/C nude mice. Co-transplantation of hepatocytes with tumor cells did not increase tumorigenesis of the tumor cells.

CONCLUSION: Immunomagnetic depletion appears to be an effective method of purging contaminating tumor cells to below threshold for likely tumorigenesis. Along with improved techniques for isolation of large numbers of viable hepatocytes, normal liver resected for neoplasia has potential as another clinically useful source of hepatocytes for transplantation.

Keywords: Hepatocyte transplantation, Immunomagnetic purging, Isolation of hepatocytes