In Vitro Expansion of Human Hepatocytes Is Restricted by Telomere-Dependent Replicative Aging

Henning Wege, Michael S. Chui, Hai T. Le, Stephen C. Strom, Mark A Zern

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Currently, different techniques to expand human hepatocytes in vitro are being investigated to generate enough cells for liver-directed cell therapies. However, based on observations in fibroblasts and other cell types, telomere attrition limits the proliferative capacity of normal somatic cells. Therefore, we explored whether telomere-dependent replicative aging restricts the in vitro proliferation of human hepatocytes. Subpopulations of cells isolated from a neonatal liver and characterized as hepatocyte derived by RT-PCR and flow cytometry started to proliferate 5-7 days after plating and were termed proliferating human hepatocytes (PHH). Following retroviral-mediated transduction of the catalytic telomerase subunit, telomerase reverse transcriptase (hTERT), telomerase activity increased from almost undetectable levels to levels as high as in HepG2 and other telomerase-positive cell lines. As expected, untransduced PHH progressively lost telomeric repeats and arrested after 30-35 cell divisions with telomeres of less than 5 kilo bases. In comparison, telomerase-reconstituted PHH maintained elongated telomeres and continued to proliferate as shown by colorimetric assays and cell counts. In this study, telomere stabilization extended the proliferative capacity of in vitro proliferating human neonatal hepatocytes. Therefore, telomere attrition needs to be addressed when developing techniques to expand human hepatocytes.

Original languageEnglish (US)
Pages (from-to)897-906
Number of pages10
JournalCell Transplantation
Volume12
Issue number8
StatePublished - 2004

Fingerprint

Telomere
Hepatocytes
Aging of materials
Telomerase
Cells
Liver
Flow cytometry
In Vitro Techniques
Fibroblasts
Cell- and Tissue-Based Therapy
Plating
Cell Division
Assays
Flow Cytometry
Stabilization
Cell Count
Cell Line
Polymerase Chain Reaction

Keywords

  • Hepatocyte
  • Immortality
  • Proliferation
  • Senescence
  • Telomerase

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation

Cite this

Wege, H., Chui, M. S., Le, H. T., Strom, S. C., & Zern, M. A. (2004). In Vitro Expansion of Human Hepatocytes Is Restricted by Telomere-Dependent Replicative Aging. Cell Transplantation, 12(8), 897-906.

In Vitro Expansion of Human Hepatocytes Is Restricted by Telomere-Dependent Replicative Aging. / Wege, Henning; Chui, Michael S.; Le, Hai T.; Strom, Stephen C.; Zern, Mark A.

In: Cell Transplantation, Vol. 12, No. 8, 2004, p. 897-906.

Research output: Contribution to journalArticle

Wege, H, Chui, MS, Le, HT, Strom, SC & Zern, MA 2004, 'In Vitro Expansion of Human Hepatocytes Is Restricted by Telomere-Dependent Replicative Aging', Cell Transplantation, vol. 12, no. 8, pp. 897-906.
Wege, Henning ; Chui, Michael S. ; Le, Hai T. ; Strom, Stephen C. ; Zern, Mark A. / In Vitro Expansion of Human Hepatocytes Is Restricted by Telomere-Dependent Replicative Aging. In: Cell Transplantation. 2004 ; Vol. 12, No. 8. pp. 897-906.
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