Establishment of immortalized human hepatic stellate scavenger cells to develop bioartificial livers

Transplantation. 2003 Jun 15;75(11):1873-80. doi: 10.1097/01.TP.0000064621.50907.A6.


Background: Maintenance of liver-specific functions has been shown to be stabilized by co-cultivation of hepatocytes with hepatic stellate cells (HSC). Because the limited lifespan of human HSC is a major hurdle to their use, the authors report here the amplification of human HSC populations in vitro by retroviral transfer of human telomerase reverse transcriptase (hTERT).

Methods: Human HSC strain LI 90 cells were transduced with a retroviral vector SSR#197 expressing hTERT and green fluorescent protein (GFP) cDNA flanked by a pair of loxP. TWNT-1, one of SSR#197-immortalized HSC, was characterized. Differentiated liver functions were evaluated in an immortalized human hepatocyte NKNT-3-TWNT-1 co-culture system.

Results: TWNT-1 cells showed differential functions of HSC, including uptake of acetylated low-density lipoproteins and synthesis of collagen type I and hepatocyte growth factor. Efficient excision of the retrovirally transferred hTERT and GFP cDNAs was achieved by TAT-mediated expression of the Cre recombinase and subsequent GFP-negative cell sorting. When co-cultured with TWNT-1 cells, NKNT-3 increased protein expression of the detoxifying cytochrome P450-associated protein isoenzymes 3A4 and 2C9 and urea synthesis.

Conclusions: TWNT-1 cells could be valuable in the study of integrated liver functions and contribute to the optimization of liver cell therapies and bioartificial livers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Actins / genetics
  • Animals
  • Aryl Hydrocarbon Hydroxylases / genetics
  • Carbocyanines
  • Cell Division
  • Cell Line, Transformed
  • Cholesterol, LDL / pharmacokinetics
  • Coculture Techniques
  • Cytochrome P-450 CYP2C9
  • Cytochrome P-450 CYP3A
  • Cytochrome P-450 Enzyme System / genetics
  • DNA-Binding Proteins
  • Endocytosis
  • Female
  • Fluorescent Dyes
  • Gene Expression
  • Gene Products, tat / genetics
  • Green Fluorescent Proteins
  • Hepatocytes / cytology*
  • Humans
  • Indicators and Reagents / metabolism
  • Integrases / genetics
  • Liver, Artificial*
  • Luminescent Proteins / genetics
  • Mice
  • Mice, SCID
  • Middle Aged
  • Receptor, Platelet-Derived Growth Factor beta / genetics
  • Telomerase / genetics
  • Telomerase / metabolism
  • Telomere / metabolism
  • Urea / metabolism
  • Viral Proteins / genetics
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism


  • 3,3'-dihexadecylindocarbocyanine
  • Actins
  • Carbocyanines
  • Cholesterol, LDL
  • DNA-Binding Proteins
  • Fluorescent Dyes
  • Gene Products, tat
  • Indicators and Reagents
  • Luminescent Proteins
  • Viral Proteins
  • Green Fluorescent Proteins
  • Urea
  • Cytochrome P-450 Enzyme System
  • CYP2C9 protein, human
  • Cytochrome P-450 CYP2C9
  • Aryl Hydrocarbon Hydroxylases
  • CYP3A protein, human
  • Cytochrome P-450 CYP3A
  • Receptor, Platelet-Derived Growth Factor beta
  • Cre recombinase
  • Integrases
  • Telomerase
  • beta-Galactosidase