title: Manipulating the Proliferative Potential of Cardiomyocytes by Gene Transfer.
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.
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano, 99, Trieste, 34149, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Prosdocimo
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano, 99, Trieste, 34149, Italy. giacca@icgeb.org.|Department of Medical, Surgical and Health Sciences, University of Trieste, Trieste, Italy. giacca@icgeb.org.
authors.forename: Mauro
authors.initials: M
authors.lastname: Giacca
mesh_terms: D000818:Animals; D064107:Aurora Kinase B / Q000378:metabolism; D015415:Biomarkers; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics; D005260:Female; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D006657:Histones / Q000378:metabolism; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics; D008856:Microscopy, Fluorescence; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D014161:Transduction, Genetic; D014162:Transfection; D019076:Transgenes
chemical_list: D015415:Biomarkers; D006657:Histones; D035683:MicroRNAs; D064107:Aurora Kinase B
keywords: AAV vectors; Aurora B; BrdU; Cardiomyocytes; EdU; MicroRNAs; Phospho-histone 3