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


CONCLUSIONS
HMAs and HDACs enhanced in vitro differentiation of MSCs, which was maximized when the two drugs were combined, with HMA having the dominant effect.
Laboratory of Hematological Disease and Transplant Immunology, Seoul, Republic of Korea.
Hye Joung
HJ
Kim
Laboratory of Hematological Disease and Transplant Immunology, Seoul, Republic of Korea.
Yong-Rim
YR
Kwon
Laboratory of Hematological Disease and Transplant Immunology, Seoul, Republic of Korea.
Yu-Jin
YJ
Bae
Laboratory of Hematological Disease and Transplant Immunology, Seoul, Republic of Korea. yoojink@catholic.ac.kr.|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. yoojink@catholic.ac.kr.
Yoo-Jin
YJ
Kim
D001374:Azacitidine / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects; D004359:Drug Therapy, Combination; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D055495:Neurogenesis / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects
D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C012589:trichostatin A; D001374:Azacitidine
5-Azacytidine; Differentiation; Histone deacetylase inhibitor; Hypomethylating agents; Mesenchymal stem cells; Trichostatin