Hanyang Med Rev.  2014 Nov;34(4):165-172. 10.7599/hmr.2014.34.4.165.

Hepatocyte Isolation, Culture, and Its Clinical Applications

• Lee DH
• Lee KW

• KMID: 2168348
• DOI: http://doi.org/10.7599/hmr.2014.34.4.165

Abstract

Hepatocytes, parenchymal cells of the liver, are specially differentiated cells to perform most of the body metabolisms. Many clinicians are interested in utilizing hepatocytes as cell therapeutics. A great number of investigators have been harvesting hepatocytes using two-step portal vein perfusion method, in which Ca2+-free EDTA-containing buffer and Ca2+-enriched collagenase solution are pumped into liver in sequence. Among various attempts for long-term culture of hepatocytes, collagen gel sandwich configuration is recognized to be the most effective technique. In the biomedical field, hepatocytes have been used in three methods of applications. First is hepatocyte transplantation for the treatment of acute, chronic liver failure and metabolic diseases. Donated livers not suitable for organ transplantation are rare, which is the major human hepatocyte source. This shortage of human hepatocyte source is expected to be resolved by virtue of rapid progressing stem cell technologies. The second application is biological components of bioartificial liver (BAL) system for acute liver failure patients. Due to the lack of functional activity of clinically studied BAL systems and difficulty of establishing a manufacturing system for ready-to-use, additional research activities are stagnated. The third utilization of hepatocytes is in vitro drug screening studies such as drug metabolism, transport, biliary excretion, and toxicity tests. If cell therapeutic treatments using hepatocytes are clinically valuable to some types of liver diseases, the demand for liver transplantation would be significantly diminished.

Keyword

Hepatocyte Isolation; Hepatocyte Transplantation; Liver, Artificial

MeSH Terms

Collagen
Collagenases
Drug Evaluation, Preclinical
End Stage Liver Disease
Hepatocytes*
Humans
Liver
Liver Diseases
Liver Failure, Acute
Liver Transplantation
Liver, Artificial
Metabolic Diseases
Metabolism
Organ Transplantation
Perfusion
Portal Vein
Research Personnel
Stem Cells
Toxicity Tests
Transplants
Virtues
Collagen
Collagenases

Figure

• 

  Fig. 1 Brief history of hepatocyte isolation techniques. (A) mechanical dissociation, (B) Ca2+ chelating, (C) enzyme digestion of tissue fragments, (D) two-step enzyme perfusion via portal vein.

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  Fig. 2 Schematic diagram of culture configurations of hepatocytes using ECM components. Arrow roughly indicates progress direction of culture techniques. (A) dry coated with collagen or fibronectin, (B) hydrogel of collagen, (C) floating collagen gel, (D) laminin-rich gel (Matrigel), (E) sandwiched by two layers of collagen gel, (F) membrane-supported sandwich culture.

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