Abstract
In contrast to prenatal life, cardiomyocyte proliferation in mammals is rapidly blunted after birth; as a consequence, clinically significant cardiac regeneration does not occur in adulthood. Thus, the modulation of cardiomyocyte proliferation by gene transfer offers an invaluable opportunity to both understand the mechanisms regulating renewal of these cells in the fetus and identify novel strategies for myocardial repair.
In this Chapter, we report an exhaustive protocol to isolate, culture, and manipulate the properties of neonatal ventricular rat cardiomyocytes by small RNA transfection or transduction with viral vectors based on the adeno-associated virus, which exhibit exquisite tropism for these cells. We also provide techniques to assess DNA synthesis and cell proliferation.
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Acknowledgments
This work was supported by Grant 14CVD04 from the Leducq Foundation Transatlantic Network of Excellence, projects FIRB RBAP11Z4Z9 and PRIN 2010RNXM9C from the Ministry of Education, University and Research (MIUR), Italy.
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Prosdocimo, G., Giacca, M. (2017). Manipulating the Proliferative Potential of Cardiomyocytes by Gene Transfer. In: Di Nardo, P., Dhingra, S., Singla, D. (eds) Adult Stem Cells. Methods in Molecular Biology, vol 1553. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6756-8_4
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DOI: https://doi.org/10.1007/978-1-4939-6756-8_4
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