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Updated: May 6, 2026

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure
Published on: September 3, 2013
Hepatocyte isolation from unused/rejected livers for transplantation: initial step toward hepatocyte transplantation,
Mahdokht Hossein Aghdaie1, Bita Geramizadeh, Negar Azarpira
1Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, IR Iran.
Background:
Hepatocyte transplantation is being used in patients with liver-based metabolic disorders and acute liver failure. Hepatocytes can be isolated from unused/rejected livers under sterile conditions.
Objectives:
The quality of the hepatocytes is very important and the main and initial step in hepatocyte transplantation is hepatocyte isolation. In this study we tried to set up the methods of hepatocyte isolation in order to use the high quality cells in acute liver failure or congenital metabolic disorders.
Materials And Methods:
In this study, during a year, hepatocytes were isolated from 7 unused/rejected livers among more than 300 harvested livers in Shiraz University of Medical Sciences. The two step collagenase perfusion method was used under GMP (Good manufacturing practice) for hepatocyte isolation.
Results:
Highly quality hepatocytes with high viability and low contamination were isolated. The mean viability was 71.8% ± 21.7. In the first 4 cases microbial contamination by Staphylococci, Diphtheroid and Klebsiella was detected, however the last 3 cases were free of any micro organisms. After 5 weeks of cryopreservation in -140°C, the cell viability was still acceptable.
Conclusions:
Hepatocyte isolation can be performed as the main and initial step for cell transplantation from unused/rejected liver. It is the first experience in Iran.
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