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Enhancement of human mesenchymal stem cell differentiation by combination treatment with 5-azacytidine and trichostatin A

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Abstract

Objective

To enhance the differentiation of mesenchymal stem cells (MSCs) and their epigenetic status by modification using hypomethylating agents (HMAs) and histone deacetylase inhibitors (HDACs).

Results

Treatment with 5-azacytidine or 5-azacytidine plus trichostatin A (TSA) increased expression of Runx-2, BDNF and Sox-9 compared with the control or TSA groups. Maximal increases of 4.1-, 4.5-, and 8.3-fold in Runx-2, BDNF, and Sox-9 transcript levels, respectively, were observed in the 5-azacytidine plus TSA group. Similar to the expression pattern of key regulatory molecules, differentiation to each lineage was also enhanced considerably in the 5-azacytidine or in the 5-azacytidine plus TSA groups. Quantitative analyses at the protein level showed 8.9-, 26.8-, 27.9-, and 28.5-fold upregulation of osterix, MAP-2, nestin, and type II collagen), respectively.

Conclusion

HMAs and HDACs enhanced in vitro differentiation of MSCs, which was maximized when the two drugs were combined, with HMA having the dominant effect.

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Acknowledgments

This study was supported by a grant from Sanofi Aventis, Korea. This research was partly supported by the National Research Foundation (NRF) funded by the Korean government (No. NRF-2012R1A1A2043735).

Supporting information

Supplementary Table 1—Primers used for real time-PCR analysis.

Supplementary Fig. 1—Flow cytometric analysis of the MSCs.

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Authors and Affiliations

  1. Laboratory of Hematological Disease and Transplant Immunology, Seoul, Republic of Korea

    Hye Joung Kim, Yong-Rim Kwon, Yu-Jin Bae & Yoo-Jin Kim

  2. Department of Hematology, Seoul St. Mary’s Hospital, College of Medicine, Catholic Blood and Marrow Transplantation Center, Catholic University Of Korea, 505 Banpo-Dong, Seocho-Gu, Seoul, 137-701, Republic of Korea

    Yoo-Jin Kim

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  1. Hye Joung Kim
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  2. Yong-Rim Kwon
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Corresponding author

Correspondence to Yoo-Jin Kim.

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Kim, H.J., Kwon, YR., Bae, YJ. et al. Enhancement of human mesenchymal stem cell differentiation by combination treatment with 5-azacytidine and trichostatin A. Biotechnol Lett 38, 167–174 (2016). https://doi.org/10.1007/s10529-015-1949-3

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  • DOI: https://doi.org/10.1007/s10529-015-1949-3

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