Summary
The pathogenesis of heart disease and the development of myocardial failure are highly dependent upon the cardiomyocyte—the basic contractile cell within the heart. Understanding and elucidating the complex networks that regulate cardiomyocyte function are central to the development of specific target-based therapeutic interventions. The relative recent advances in molecular genetics and generation and routine usage of transgenic and knockout mouse models have further necessitated that previously established cardiomyocyte methods be adapted for the isolation, culture, and study of primary adult murine cardiomyocytes, both freshly isolated and in culture. Such adaptation is based not only upon scalability of established techniques, as the mouse heart is an order of magnitude smaller than the rat heart, but also upon properties unique to the mouse. This chapter therefore describes current methods for the isolation, culture, and functional analysis of adult murine cardiomyocytes.
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References
American Heart Association. (2005) Heart disease and stroke statistics – 2005 update. American Heart Association. Dallas, Texas.
Braunwald, E., and Bristow, M. R. (2000) Congestive heart failure: fifty years of progress. Circulation 102, 14–23.
Zhou, Y. Y., Wang, S. Q., Zhu, W. Z., Chruscinski, A., Kobilka, B. K., Ziman, B., Wang, S., Lakatta, E. G., Cheng, H., and Xiao, R. P. (2000) Culture and adenoviral infection of adult mouse cardiac myocytes: methods for cellular genetic physiology. Am. J. Physiol. Heart Circ. Physiol. 279, H429–H436.
O’Connell, T. D., Ni, Y. G., Lin, K.-M., Han, H., and Yan, Z. (2003) Isolation and culture of adult mouse cardiac myocytes for signaling studies. AFCS Research Reports [online] 1(5), http://www.signaling-gateway.org/reports/v1/CM0005/CM0005.htm.
Jain, M., Lim, C. C., Nagata, K., Davis, V. M., Milstone, D. S., Liao, R., and Mortensen, R. M. (2001) Targeted inactivation of Galpha(i) does not alter cardiac function or beta-adrenergic sensitivity. Am. J. Physiol. Heart Circ. Physiol. 280, H569–H575.
Lim, C. C., Apstein, C. S., Colucci, W. S., and Liao, R. (2000) Impaired cell shortening and relengthening with increased pacing frequency are intrinsic to the senescent mouse cardiomyocyte. J. Mol. Cell. Cardiol. 32, 2075–2082.
Michael, A., Haq, S., Chen, X., Hsich, E., Cui, L., Walters, B., Shao, Z., Bhattacharya, K., Kilter, H., Huggins, G., Andreucci, M., Periasamy, M., Solomon, R. N., Liao, R., Patten, R., Molkentin, J. D., and Force, T. (2004) Glycogen synthase kinase-3beta regulates growth, calcium homeostasis, and diastolic function in the heart. J. Biol. Chem. 279, 21383–21393.
Matsui, T., Li, L., Wu, J. C., Cook, S. A., Nagoshi, T., Picard, M. H., Liao, R., and Rosenzweig, A. (2002) Phenotypic spectrum caused by transgenic overexpression of activated Akt in the heart. J. Biol. Chem. 277, 22896–22901.
Nagata, K., Ye, C., Jain, M., Milstone, D. S., Liao, R., and Mortensen, R. M. (2000) Galpha(i2) but not Galpha(i3) is required for muscarinic inhibition of contractility and calcium currents in adult cardiomyocytes. Circ. Res. 87, 903–909.
Acknowledgments
The authors thank all members of Cardiac Muscle Research Laboratory at Brigham and Women’s Hospital, Harvard Medical School, and, in particular, Drs Lei Cui and Bo Wang for their technical expertise. This work was supported by National Institutes of Health grants HL-73756, HL-67297, and HL-71775 (RL).
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Liao, R., Jain, M. (2007). Isolation, Culture, and Functional Analysis of Adult Mouse Cardiomyocytes. In: Sreejayan, N., Ren, J. (eds) Vascular Biology Protocols. Methods in Molecular Medicine™, vol 139. Humana Press. https://doi.org/10.1007/978-1-59745-571-8_16
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DOI: https://doi.org/10.1007/978-1-59745-571-8_16
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