5-Azacytidine facilitates osteogenic gene expression and differentiation of mesenchymal stem cells by alteration in DNA methylation.
Mesenchymal stem cells (MSCs) are considered to be one of the most promising therapeutic cell sources as they encompass a plasticity of multiple cell lineages. The challenge in using these cells lies in developing well-defined protocols for directing cellular differentiation to generate a desired lineage. In this study, we investigated the effect of 5-azacytidine, a DNA demethylating agent, on osteogenic differentiation of MSCs. The cells were exposed to 5-azacytidine in culture medium for 24 h prior to osteogenic induction. Osteogenic differentiation was determined by several the appearance of a number of osteogenesis characteristics, including gene expression, ALP activity, and calcium mineralization. Pretreatment of MSCs with 5-azacytidine significantly facilitated osteogenic differentiation and was accompanied by hypomethylation of genomic DNA and increased osteogenic gene expression. Taking dlx5 as a representative, methylation alterations of the "CpG island shore" in the promoter caused by 5-azacytidine appeared to contribute to osteogenic differentiation.
Zhou
Guo-Shun
GS
Key Laboratory of Molecular Medicine, Huzhou Central Hospital, 313100, Huzhou, Zhejiang, People's Republic of China. gfz@mail.huptt.zj.cn.
Zhang
Xiao-Lei
XL
College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China.|Key Lab for Cell and Gene Engineering of Zhejiang Province, Hangzhou, People's Republic of China.
Wu
Jun-Ping
JP
College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China.|Key Lab for Cell and Gene Engineering of Zhejiang Province, Hangzhou, People's Republic of China.
Zhang
Rui-Peng
RP
College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China.|Key Lab for Cell and Gene Engineering of Zhejiang Province, Hangzhou, People's Republic of China.
Xiang
Li-Xin
LX
College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China.|Key Lab for Cell and Gene Engineering of Zhejiang Province, Hangzhou, People's Republic of China.
Dai
Li-Cheng
LC
Key Laboratory of Molecular Medicine, Huzhou Central Hospital, 313100, Huzhou, Zhejiang, People's Republic of China.
Shao
Jian-Zhong
JZ
College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China. shaojz@zju.edu.cn.|Key Lab for Cell and Gene Engineering of Zhejiang Province, Hangzhou, People's Republic of China. shaojz@zju.edu.cn.