Telomerase-mediated life-span extension of human primary fibroblasts by human artificial chromosome (HAC) vector

Telomerase-mediated life-span extension enables the expansion of normal cells without malignant transformation, and thus has been thought to be useful in cell therapies. Currently, integrating vectors including the retrovirus are used for human telomerase reverse transcriptase (hTERT)-mediated expan...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2008-05, Vol.369 (3), p.807-811
Main Authors: Shitara, Shingo, Kakeda, Minoru, Nagata, Keiko, Hiratsuka, Masaharu, Sano, Akiko, Osawa, Kanako, Okazaki, Akiyo, Katoh, Motonobu, Kazuki, Yasuhiro, Oshimura, Mitsuo, Tomizuka, Kazuma
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
Animals
Cell Division
CELL PROLIFERATION
Cellular proliferation
Cellular Senescence
CHO Cells
CHROMOSOMES
Chromosomes, Artificial, Human - genetics
Cricetinae
Cricetulus
FIBROBLASTS
Fibroblasts - enzymology
Fibroblasts - physiology
GALACTOSIDASE
Gene Transfer Techniques
Genetic Vectors - genetics
hTERT
Human artificial chromosome (HAC)
Humans
LIFE SPAN
Life-span extension
MUTAGENESIS
Non-integrating vector
ONCOGENES
Primary cultured cells
Telomerase
Telomerase - analysis
Telomerase - genetics
THERAPY
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Summary:Telomerase-mediated life-span extension enables the expansion of normal cells without malignant transformation, and thus has been thought to be useful in cell therapies. Currently, integrating vectors including the retrovirus are used for human telomerase reverse transcriptase (hTERT)-mediated expansion of normal cells; however, the use of these vectors potentially causes unexpected insertional mutagenesis and/or activation of oncogenes. Here, we established normal human fibroblast (hPF) clones retaining non-integrating human artificial chromosome (HAC) vectors harboring the hTERT expression cassette. In hTERT-HAC/hPF clones, we observed the telomerase activity and the suppression of senescent-associated SA-β-galactosidase activity. Furthermore, the hTERT-HAC/hPF clones continued growing beyond 120 days after cloning, whereas the hPF clones retaining the silent hTERT-HAC senesced within 70 days. Thus, hTERT-HAC-mediated episomal expression of hTERT allows the extension of the life-span of human primary cells, implying that gene delivery by non-integrating HAC vectors can be used to control cellular proliferative capacity of primary cultured cells.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2008.02.119