Protocol

Human Cell Culture Protocols

Volume 806 of the series Methods in Molecular Biology pp 99-120

Date:

Human Hepatocytes: Isolation, Culture, and Quality Procedures

  • Daniel KnobelochAffiliated withCytonet Gmbh
  • , Sabrina EhnertAffiliated withDepartment of Traumatology, Eberherd karls Universität Tübingen
  • , Lilianna SchyschkaAffiliated withDepartment of Traumatology, MRI, Technische Universität München
  • , Peter BüchlerAffiliated withDepartment of General Surgery, TU Munich, MRI
  • , Michael SchoenbergAffiliated withDepartment of Gastroenterology & Hepatology, Medical University Hospital
  • , Jörg KleeffAffiliated withDepartment of General Surgery, TU Munich, MRI
  • , Wolfgang E. ThaslerAffiliated withDepartment of Surgery, Ludwigs-Maximillian Universität Münchern
  • , Natascha C. NusslerAffiliated withDepartment of General and Visceral Surgery, Klinikum Neuperlach
  • , Patricio GodoyAffiliated withIFADO, Leibniz Research Centre for Working Environment and Human Factors, Dortmund University
    • , Jan HengstlerAffiliated withIFADO, Leibniz Research Centre for Working Environment and Human Factors, Dortmund University
    • , Andreas K. NusslerAffiliated withDepartment of Traumatology, Eberherd karls Universität Tübingen Email author 

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Abstract

The use of isolated human liver cells in research and development has gained increasing interest during the past years. The possible application may vary between elucidation of new biochemical pathways in liver diseases, drug development, safety issues, and new therapeutic strategies up to direct clinical translation for liver support. However, the isolation of human liver cells requires a well-developed logistic network among surgeons, biologists, and technicians to obtain a high quality of cells. Our laboratories have been involved in various applications of human liver cells and we have long-lasting experiences in human liver cell isolation and their application in R&D. We here summarize the present protocol of our laboratories for cell isolation from normal resected liver tissue, the most common tissue available. In addition, we discuss the necessary network in the clinic and quality controls to maintain human liver cells in culture and the effect of 3D extracellular matrix in cultured cells which results in preservation of hepatocyte epithelial polarity in the form of bile canaliculi and repression of epithelial to mesenchymal transitions occurring in 2D cultures.

Key words

Human hepatocytes Cell isolation Hepatocyte culture Quality control Metabolic competence