title: Combined use of N-acetylcysteine and Liberase improves the viability and metabolic function of human hepatocytes isolated from human liver.
abstract: OBJECTIVE
  Successful hepatocyte isolation is critical for continued development of cellular transplantation. However, most tissue available for research is from diseased liver, and the results of hepatocyte isolation from such tissue are inferior compared with normal tissue. Liberase and N-acetylcysteine (NAC) have been shown separately to improve viability of isolated hepatocytes. This study aims to determine the effect of Liberase and NAC in combination on human hepatocyte isolation from normal and diseased liver tissues.
  
  
  METHODS
  Hepatocytes were isolated from 30 liver specimens through the use of a standard collagenase digestion technique (original protocol) and another 30 with the addition of NAC and standard collagenase substituted by Liberase (new protocol). Viability and success, defined as maintenance of cell adhesion and morphology for 48 hours, were assessed. Metabolic function was assessed by means of albumin and urea synthesis.
  
  
  RESULTS
  Baseline factors were similar for both groups. The delay to tissue processing was slightly shorter in the new protocol group (median, 2 versus 4 hours; P = 0.007). The success rate improved from 12 of 30 (40.0%) to 21 of 30 (70.0%) with the use of the new protocol (P = 0.037), and median viable cell yield increased from 7.3 × 10(4) to 28.3 × 10(4) cells/g tissue (P = 0.003). After adjusting for delay, success rate (P = 0.014) and viable cell yield/g tissue (P = 0.001) remained significantly improved. Albumin and urea synthesis were similar or superior in the new protocol group.
  
  
  CONCLUSIONS
  NAC and Liberase improve the success of hepatocyte isolation, with a significantly higher yield of viable cells. The use of these agents may improve the availability of hepatocytes for transplantation and laboratory research.
authors.affiliation: National Institute for Health Research Biomedical Unit and Centre for Liver Research, University of Birmingham, Birmingham, United Kingdom. Electronic address: davidbartlett1@nhs.net.
authors.forename: David C
authors.initials: DC
authors.lastname: Bartlett
authors.affiliation: National Institute for Health Research Biomedical Unit and Centre for Liver Research, University of Birmingham, Birmingham, United Kingdom.
authors.forename: James
authors.initials: J
authors.lastname: Hodson
authors.affiliation: National Institute for Health Research Biomedical Unit and Centre for Liver Research, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Ricky H
authors.initials: RH
authors.lastname: Bhogal
authors.affiliation: National Institute for Health Research Biomedical Unit and Centre for Liver Research, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Janine
authors.initials: J
authors.lastname: Youster
authors.affiliation: National Institute for Health Research Biomedical Unit and Centre for Liver Research, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Phil N
authors.initials: PN
authors.lastname: Newsome
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects*; D017364:Collagenases / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007515:Islets of Langerhans / Q000187:drug effects / Q000473:pathology; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D013820:Thermolysin / Q000494:pharmacology*; D014180:Transplantation / Q000379:methods
chemical_list: D017364:Collagenases; C106635:Liberase; D013820:Thermolysin; D000111:Acetylcysteine
keywords: Liberase; acetylcysteine; cell transplantation; hepatocytes