Abstract
Purpose: Bone marrow mesenchymal stem cells (BMSCs) are pluripotent and thought to be an important source for tissue engineering. Normal human BMSCs entered into senescence around PDL 30. It would be attractive to immortalize human BMSCs, while maintaining adequate qualities. Here we present a potentially novel strategy for establishing immortalized human BMSC lines using retrovirus-mediated gene transfer of human telomerase reverse transcriptase (hTERT). Methods: Normal human BMSCs were transduced with a retroviral vector SSR#197 encoding hTERT and green fluorescent protein (GFP) cDNAs. After retroviral transduction, human BMSCs were subjected to flow cytometric cell sorting for obtaining GFP-positive clones. Growth property, telomerase activity and differentiation potentials were investigated in the clones. Results: The expression of CD44 was positive in all clones. Highly proliferative clones showed the expression of CD34 and SSEA, and less proliferative clones revealed high expression of CD44. The gene expression profile was different among the 20 clones established. Clone No. 12, YKNK-12, showed low proliferation ability and high potential of differentiating into adipocytes, osteoblasts and chondrocytes. Conclusion: We have demonstrated successful retroviral transduction of human BMSCs with hTERT and suggested the presence of heterogeneity of BMSC clones.
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