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Characterization of a growth-elevated cell line of human bone marrow-derived mesenchymal stem cells by SV40 T-antigen

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Abstract

Human bone marrow-derived mesenchymal stem cells (hMSCs) are capable of self-renewal and differentiation into various tissue lineages, attracting attention as tools for use in cell therapy. However, hMSCs have very poor proliferative capacity and a short life span in culture. To overcome this problem, we expressed the T antigen of SV40 in hMSCs because it is known to have the ability to elevate the growth rate of various primary animal cells. We obtained several hMSCs lines (hMSCs-T) known for stable expression of T antigen. Cells expressing T antigen proliferated on the monolayer of hMSCs, forming high density foci. hMSCs-T showed changed morphology and improved growth rate and life span, and demonstrated preservation of the potential for differentiation into osteoblasts. In addition, hMSCs-T did not proliferate in soft agar culture, indicating that the cells did not transform into tumor cells. In order to evaluate metabolic change of amino acids in hMSCs-T compared to primary hMSCs, we investigated altered amino acids (AA) with gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode (GC-SIM-MS). A total of 14 AAs were positively measured. Results from the Student’s t-test on the hMSCs group mean of the hMSCs-T group showed significantly elevated levels of glycine, proline, pipecolic acid, aspartic acid, lysine and tryptophan, whereas valine, leucine and isoleucine as branched-chain amino acids (BCAAs), and phenylalanine showed a significant decrease. Altered AAs metabolic pattern in the hMSCs-T may explain the disturbance of AA metabolism related to the expression of SV40 T antigen in hMSCs.

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Authors and Affiliations

  1. Department of Microbiology, College of Natural Sciences, Changwon National University, Changwon, 641-773, Korea

    Kyung Soo Lee & Dong Wan Kim

  2. Binex Co., Ltd. Sahagu, Busan, 604-846, Korea

    Kyung Soo Lee

  3. Department of Physiology and Department of Biomedical Sciences, Ajou University School of Medicine, Suwon, 443-721, Korea

    Jeom Soon Shim & Gwang Lee

  4. College of Pharmacy, Sunchon National University, Suncheon, 540-742, Korea

    Man Joeng Paik

  5. Department of Biology, College of Natural Sciences, Changwon National University, Changwon, 641-773, Korea

    Woo Hong Joo

  6. Department of Biochemistry, Pusan National University School of Medicine, Yangsan, 626-870, Korea

    Sun Hee Kim

Authors
  1. Kyung Soo Lee
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  2. Jeom Soon Shim
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  3. Man Joeng Paik
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  4. Woo Hong Joo
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  5. Sun Hee Kim
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  6. Gwang Lee
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  7. Dong Wan Kim
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Correspondence to Gwang Lee or Dong Wan Kim.

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These authors contributed equally to this work

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Lee, K.S., Shim, J.S., Paik, M.J. et al. Characterization of a growth-elevated cell line of human bone marrow-derived mesenchymal stem cells by SV40 T-antigen. Biotechnol Bioproc E 20, 498–505 (2015). https://doi.org/10.1007/s12257-014-0730-0

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  • DOI: https://doi.org/10.1007/s12257-014-0730-0

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