Skip to main content

Cardiomyocyte Preparation, Culture, and Gene Transfer

  • Protocol
Cardiac Gene Expression

Part of the book series: Methods in Molecular Biology ((MIMB,volume 366))

Abstract

Neonatal rat ventricular myocytes (NRVMs) cultured in vitro have been used as a model system for easily recreating and studying several cardiac molecular conditions, such as hypertrophy, oxygen deprivation, and gene expression. However, low efficiency of gene transfer has often represented one of the major limitations of this technique. In this chapter we describe in detail how to isolate NRVMs from neonatal rat heart and the optimal conditions for their long-term culture. Different cardiomyocyte transfection methodologies, based on viral or viral/chemical delivery carriers, are also discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Simpson, P. and Savion, S. (1982) Differentiation of rat myocytes in single cell cultures with and without proliferating nonmyocardial cells. Cross-striations, ultrastructure, and chronotropic response to isoproterenol. Circ. Res. 50, 101–116.

    CAS  PubMed  Google Scholar 

  2. Simpson, P., McGrath, A., and Savion, S. (1982) Myocyte hypertrophy in neonatal rat heart cultures and its regulation by serum and by catecholamines. Circ. Res. 51, 787–801.

    CAS  PubMed  Google Scholar 

  3. Simpson, P. (1983) Norepinephrine-stimulated hypertrophy of cultured rat myocardial cells is an alpha 1-adrenergic response. J. Clin. Invest. 72, 732–738.

    Article  CAS  PubMed  Google Scholar 

  4. Simpson, P. (1985) Stimulation of hypertrophy of cultured neonatal rat heart cells through an alpha 1-adrenergic receptor and induction of beating through an alpha 1-and beta 1-adrenergic receptor interaction. Evidence for independent regulation of growth and beating. Circ. Res. 56, 884–894.

    CAS  PubMed  Google Scholar 

  5. Musters, R. J., Post, J. A., and Verkleij, A. J. (1991) The isolated neonatal rat-cardiomyocyte used in an in vitro model for ‘ischemia’. A morphological study. Biochem. Biophys. Acta 1091, 270–277.

    Article  CAS  PubMed  Google Scholar 

  6. Iwaki, K., Chi, S. H., Dillmann, W. H., and Mestril, R. (1993) Induction of HSP70 in cultured rat neonatal cardiomyocytes by hypoxia and metabolic stress. Circulation 87, 2023–2032.

    CAS  PubMed  Google Scholar 

  7. Tanaka, M., Ito, H., Adachi, S., et al. (1994) Hypoxia induces apoptosis with enhanced expression of Fas antigen messenger RNA in cultured neonatal rat cardiomyocytes. Circ Res. 75, 426–433.

    CAS  PubMed  Google Scholar 

  8. Shimojo, T., Hiroe, M., Ishiyama, S., Ito, H., Nishikawa, T., and Marumo F. (1999) Nitric oxide induces apoptotic death of cardiomyocytes via a cyclic-GMP-dependent pathway. Exp. Cell Res. 247, 38–47.

    Article  CAS  PubMed  Google Scholar 

  9. van der Lee, K. A., Vork, M. M., De Vries, J. E., et al. (2000) Long-chain fatty acid-induced changes in gene expression in neonatal cardiac myocytes. Lipid Res. 41, 41–47.

    Google Scholar 

  10. Liu, T., Lai, H., Wu, W., Chinn, S., and Wang, P. H. (2001) Developing a strategy to define the effects of insulin-like growth factor-1 on gene expression profile in cardiomyocytes. Circ. Res. 88, 1231–1238.

    Article  CAS  Google Scholar 

  11. Charron, F., Tsimiklis, G., Arcand, M., et al. (2001) Tissue-specific GATA factors are transcriptional effectors of the small GTPase RhoA. Genes Dev. 15, 2702–2719.

    Article  CAS  PubMed  Google Scholar 

  12. Steinhelper, M. E., Lanson, N. A. Jr., Dresdner, K. P., et al. (1990) Proliferation in vivo and in culture of differentiated adult atrial cardiomyocytes from transgenic mice. Am. J. Physiol. 259, H1826–1834.

    CAS  PubMed  Google Scholar 

  13. Claycomb, W. C., Lanson, N. A. Jr., Stallworth, B. S., et al. (1998) HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. Proc. Natl. Acad. Sci. USA 95, 2979–2984.

    Article  CAS  PubMed  Google Scholar 

  14. Antin, P. B., Mar, J. H., and Ordahl, C. P. (1988) Single cell analysis of transfected gene expression in primary heart cultures containing multiple cell types. Biotechniques 6, 640–649.

    CAS  PubMed  Google Scholar 

  15. Kass-Eisler, A., Falck-Pedersen, E., Alvira, M., et al. (1993) Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo. Proc. Natl. Acad. Sci. USA 90, 11498–11502.

    Article  CAS  PubMed  Google Scholar 

  16. Kohout, T. A., O’Brian, J. J., Gaa, S. T., Lederer W. J., and Rogers T. B. (1996) Novel adenovirus component system that transferts cultured cardiac cells with high efficiency. Circ. Res. 78, 971–977.

    CAS  PubMed  Google Scholar 

  17. Maass, A., Langer, S. L., Oberdorf-Maas s, S., Neyses, L., and Leinwand L. A. (2003) Rational promoter selection for gene transfer into cardiac cells. J. Mol. Cell. Cardiol. 35, 823–831.

    Article  CAS  PubMed  Google Scholar 

  18. Godbey, W. T., Wu, K. K., and Mikos, A. G. (1999) Tracking the intracellular path of poly(efhylenimine)/DNA complexes for gene delivery. Proc. Natl. Acad. Sci. USA 96, 5177–5181.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc.

About this protocol

Cite this protocol

Maass, A.H., Buvoli, M. (2007). Cardiomyocyte Preparation, Culture, and Gene Transfer. In: Zhang, J., Rokosh, G. (eds) Cardiac Gene Expression. Methods in Molecular Biology, vol 366. Humana Press. https://doi.org/10.1007/978-1-59745-030-0_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-030-0_18

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-352-7

  • Online ISBN: 978-1-59745-030-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics