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Molecular and Cellular Biochemistry

, Volume 349, Issue 1–2, pp 117–123 | Cite as

Icariin induces mouse embryonic stem cell differentiation into beating functional cardiomyocytes

  • Xiaodong Sun
  • Xiuwei Sun
  • Xiudong Jin
  • Xiaoli Zhang
  • Chunling Liu
  • Lei Lei
  • Lianhong Jin
  • Huiwen Liu
Article

Abstract

Icariin, the primary active component of Epimedium extracts, has recently been shown to induce cardiomyocyte differentiation of murine embryonic stem (mES) cells in vitro. However, as these cardiomyocytes were not functionally characterized, the potential application of icariin-induced cardiomyocytes in clinical practice remains unclear. Therefore, in this study, we characterized the structure and function of icariin-induced cardiomyocytes to evaluate their potential application in transplantation for cardiac failure treatment. mES cells were cultured as embryoid bodies (EBs) via the direct suspension method in the presence of icariin. The protein expression profiles and ultrastructural characteristics of mES cell-derived cardiomyocytes were then characterized by immunofluorescence and transmission electron microscopy, respectively. In addition, the expression of cardiac-specific and calcium handling genes was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Cardiomyocytes induced by icariin treatment expressed the cardiac-specific proteins myosin light chain-1v (MLC1v), atrial natriuretic polypeptide (ANP), and cardiac troponin I (cTnI). Furthermore, these cells appeared to possess myofibrils organized into mature sarcomeres that had formed A and I bands. In addition, icariin treatment upregulated the mRNA levels of MLC1v, ANP, cTnI, calsequestrin (CSQ), and sodium–calcium exchanger (NCX) in these cells. Icariin induces the differentiation of mES cells into beating cardiomyocytes with normal structure and function. Therefore, these cells may have promising applications in cardiac cell therapy or tissue engineering.

Keywords

Cell differentiation Embryonic stem cells Cardiomyocyte Icariin 

Notes

Acknowledgments

We thank Miss Qiuxia Yang (Australia) for financial support, CYAGEN (Guangzhou) Biotech Ltd. for technical support, and Medical Research Center at Mudanjiang Medical College for experimental help. This study was supported by research grants from the National Natural Science Foundation of China (30971537), the Natural Science Foundation of Heilongjiang Province (ZA2006-09), Heilongjiang Province Educational Committee for backbone teacher (1055G058), and technological project of Mudanjiang Technology Bureau (G2009s2021).

Supplementary material

11010_2010_666_MOESM1_ESM.tif (114 kb)
Supplemental figure: The protocol for differentiation of mES cells into myocardial-like cells. 1 (TIFF 113 kb)

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Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Xiaodong Sun
    • 1
    • 2
  • Xiuwei Sun
    • 3
  • Xiudong Jin
    • 2
  • Xiaoli Zhang
    • 2
  • Chunling Liu
    • 2
  • Lei Lei
    • 1
  • Lianhong Jin
    • 1
  • Huiwen Liu
    • 1
  1. 1.Department of Histology and EmbryologyHarbin Medical UniversityHarbinChina
  2. 2.Department of Histology and EmbryologyMudanjiang Medical CollegeMudangjiangChina
  3. 3.Department of OncologyThe Affiliated Tumor Hospital of Harbin Medical UniversityHarbinChina

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