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. 2016 Sep 7;6(2):512–526. doi: 10.5966/sctm.2015-0226

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© 2016 The Authors Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Combination of Rb knockdown and c‐Myc overexpression transforms MSCs. (A): Parental MSCs or MSCs with genetic modification were grown in complete medium. Morphological changes were observed after genetic modification. (B): β‐galactosidase staining for analyzing senescence showed abundant senescent cells in OeMyc and Sip53‐OeMyc. Percentage of senescent cells in MSCs with different genetic modification. ∗, p < .05; ∗∗∗, p < .005 compared with MSC. (C): Cells were continuously subcultured in complete medium, and cumulative population doublings were calculated. (D): Cells were assayed for soft agar colony formation ability. (E): Cells were assayed for sphere formation in TSM. (F): Cells in TSM were subjected to Western blotting analysis for pluripotency gene expression. ∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .005 compared with MSC. Bar = 500 μm. Abbreviations: MSC, mesenchymal stem cell; OeMyc, overexpression of c‐Myc; Rb, retinoblastoma; Sip53, silence of p53; SiRb, silence of retinoblastoma; TSM, tumor sphere medium.