Isolation of human hepatocytes by a two-step collagenase perfusion procedure


The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. As such, hepatocytes are an important model for a large variety of research questions. In particular, the use of human hepatocytes is especially important in the fields of pharmacokinetics, toxicology, liver regeneration and translational research. Thus, this method presents a modified version of a two-step collagenase perfusion procedure to isolate hepatocytes as described by Seglen (1). Previously, hepatocytes have been isolated by mechanical methods. However, enzymatic methods have been shown to be superior as hepatocytes retain their structural integrity and function after isolation. This method presented here adapts the method designed previously for rat livers to human liver pieces and results in a large yield of hepatocytes with a viability of 77±10%. The main difference in this procedure is the process of cannulization of the blood vessels. Further, the method described here can also be applied to livers from other species with comparable liver or blood vessel sizes.


Projects: A: Cellular level, B: Communication of Hepatocytes and Non-Parenchymal Liver Cells, CTUs, Show cases, Virtual Liver

J Vis Exp
J Vis Exp (79):
24th Sep 2013

Serene M L Lee, Celine Schelcher, Maresa Demmel, Maria Hauner, Wolfgang E Thasler

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[Serene Lee] [Maria Hauner] [Wolfgang E. Thasler]

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Views: 2590
  • Created: 13th Jan 2014 at 15:48

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