Nitric oxide facilitates cardiomyogenesis in mouse embryonic stem cells

Shinichi Kanno*, Peter K.M. Kim, Karim Sallam, Jing Lei, Timothy R. Billiar, Larry L. Shears

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

154 Scopus citations


Stem cell therapy holds great promise for the replacement of damaged or dysfunctional myocardium. Nitric oxide (NO) has been shown to promote embryonic stem (ES) cell differentiation in other systems. We hypothesized that NO, through NO synthase gene transfer or exogenous NO exposure, would promote the differentiation of mouse ES cells into cardiomyocytes (CM). In our study, NO treatment increased both the number and the size of beating foci in embryoid body (EB) outgrowths. Within 2 weeks, 69% of the inducible NO synthase-transduced EB displayed spontaneously beating foci, as did 45% of the NO donor-treated EB, compared with only ≈15% in controls. Cardiac-specific genes and protein expression were significantly increased in NO-treated ES. Electron microscopy and immunocytochemistry revealed that these NO-induced contracting cells exhibited characteristics consistent with CM. At day 7 in culture, troponin T was expressed in 45.6 ± 20.6% of the NO-treated ES cells but in only 9.25 ± 1.77% of control cells. Interestingly, 50.4 ± 18.4% of NO-treated ES cells were troponin T-negative and annexin V-positive. This apoptotic phenotype was seen in <1% of the control ES cells. These data strongly support our hypothesis that mouse ES cells can be accelerated to differentiate into CM by NO treatment. NO may influence cardiac differentiation by both inducing a switch toward a cardiac phenotype and inducing apoptosis in cells not committed to cardiac differentiation.

Original languageEnglish
Pages (from-to)12277-12281
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number33
StatePublished - 17 Aug 2004
Externally publishedYes


Dive into the research topics of 'Nitric oxide facilitates cardiomyogenesis in mouse embryonic stem cells'. Together they form a unique fingerprint.

Cite this