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hTERT Gene Immortalized Human Adipose-Derived Stem Cells and its Multiple Differentiations: a Preliminary Investigation

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Abstract

Human adipose-derived adult stem cells (hADSCs) can express human telomerase reverse transcriptase phenotypes under an appropriate culture condition. Because adipose tissue is abundant and easily accessible, hADSCs offer a promising source of stem cells for tissue engineering application and other cell-based therapies. However, the shortage of cells number and the difficulty to proliferate, known as the “Hayflick limit” in vitro, limit their further clinical application. Here, hADSCs were transfected with human telomerase reverse transcriptase (hTERT) gene by the lentiviral vector to prolong the lifespan of stem cells and even immortalize them. Following to this, the cellular properties and functionalities of the transfected cell lines were assayed. The results demonstrated that hADSCs had been successfully transfected with hTERT gene (hTERT-ADSCs). Then, hTERT-ADSCs were initially selected by G418 and subsequently expanded over 20 passages in vitro. Moreover, the qualitative and quantitative differentiation criteria for 20 passages of hTERT-ADSCs also demonstrated that hTERT-ADSCs could differentiate into osteogenesis, chondrogenesis, and adipogenesis phenotypes in lineage-specific differentiation media. These findings confirmed that this transfection could prolong the lifespan of hADSCs.

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Abbreviations

hADSCs:

Human adipose-derived adult stem cells

hTERT:

Human telomerase reverse transcriptase

s.c.:

Subcutaneously

ALP:

Alkaline phosphatase

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Acknowledgments

This work was supported by the Fok Ying Tung Education Foundation (132027), National Science Foundation of China (81271719), the State Key Laboratory of Fine Chemicals (KF1111), and the Fundamental Research Funds for the Central Universities (DUT11SM09/DUT12JB09) and SRF for ROCS, SEM.

Conflict of interests

The authors declare that they have no competing interests.

Author information

Authors and Affiliations

  1. Department of Oncology, First Affiliated Hospital of Dalian Medical University, Dalian, People’s Republic of China

    L. Wang, H. Zhu & X. Xu

  2. Dalian R&D Center for Stem Cell and Tissue Engineering, State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian, 116024, China

    K. Song & X. Qu

  3. Department of Orthopaedics, First Affiliated Hospital of Dalian Medical University, Dalian, 116011, People’s Republic of China

    H. Wang, M. Zhang, Y. Tang & X. Yang

Authors
  1. L. Wang
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  2. K. Song
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  3. X. Qu
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  4. H. Wang
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  5. H. Zhu
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  6. X. Xu
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  7. M. Zhang
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  8. Y. Tang
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  9. X. Yang
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Corresponding author

Correspondence to H. Wang.

Additional information

L. Wang, K. Song, X. Qu, and H. Wang contributed equally to this work.

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Wang, L., Song, K., Qu, X. et al. hTERT Gene Immortalized Human Adipose-Derived Stem Cells and its Multiple Differentiations: a Preliminary Investigation. Appl Biochem Biotechnol 169, 1546–1556 (2013). https://doi.org/10.1007/s12010-012-0019-8

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  • DOI: https://doi.org/10.1007/s12010-012-0019-8

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