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Cytotechnology

, Volume 64, Issue 2, pp 197–202 | Cite as

Electron microscopic study of mouse embryonic stem cell-derived cardiomyocytes

  • Masoumeh Fakhr TahaEmail author
  • Mojtaba Rezazadeh Valojerdi
  • Leili Hatami
  • Arash Javeri
Original Research

Abstract

Differentiation of embryonic stem cell (ESC)-derived embryoid bodies (EBs) is a heterogeneous process. ESCs can differentiate in vitro into different cell types including beating cardiomyocytes. The main aim of the present study was to develop an improved preparation method for scanning electron microscopic study of ESC-derived cardiac bundles and to investigate the fine structural characteristics of mouse ESCs-derived cardiomyocytes using electron microscopy. The mouse ESCs differentiation was induced by EBs’ development through hanging drop, suspension and plating stages. Cardiomyocytes appeared in the EBs’ outgrowth as beating clusters that grew in size and formed thick branching bundles gradually. Cardiac bundles showed cross striation even when they were observed under an inverted microscope. They showed a positive immunostaining for cardiac troponin I and α-actinin. Transmission and scanning electron microscopy (TEM & SEM) were used to study the structural characteristics of ESC-derived cardiomyocytes. Three weeks after plating, differentiated EBs showed a superficial layer of compact fibrous ECM that made detailed observation of cardiac bundles impossible. We tried several preparation methods to remove unwanted cells and fibers, and finally we revealed the branching bundles of cardiomyocytes. In TEM study, most cardiomyocytes showed parallel arrays of myofibrils with a mature sarcomeric organization marked by H-bands, M-lines and numerous T-tubules. Cardiomyocytes were connected to each other by intercalated discs composed of numerous gap junctions and fascia adherences.

Keywords

Mouse embryonic stem cell Cardiomyocyte Scanning electron microscopy Ultrastructure 

Notes

Acknowledgments

This study was supported by research grants from Jehad Daneshgahi (137-1-5). We thank Dr. Hossein Baharvand for providing ESCs, Mrs. Ilnaz Sadeghi Sadr for assistance in the SEM imaging and Abbas Piriaii and Shahram Pour Beiranvand for technical assistance in the TEM study.

Supplementary material

Video: Contracting bundles of cardiomyocytes at day-14 after plating (WMV 4013 kb)

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Masoumeh Fakhr Taha
    • 1
    Email author
  • Mojtaba Rezazadeh Valojerdi
    • 2
    • 3
  • Leili Hatami
    • 4
  • Arash Javeri
    • 1
  1. 1.Department of Medical BiotechnologyNational Institute of Genetic Engineering and Biotechnology (NIGEB)TehranIran
  2. 2.Department of Anatomy, School of Medical SciencesTarbiat Modares UniversityTehranIran
  3. 3.Department of EmbryologyRoyan InstituteTehranIran
  4. 4.Department of Anatomy, School of Medical SciencesShahroud University of Medical SciencesShahroudIran

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