Biolistic transfection of freshly isolated adult ventricular myocytes.

David F Steele, Ying Dou, David Fedida

Department of Anesthesiology, Pharmacology and Therapeutics, University of British Columbia, Vancouver, BC, Canada.

Methods in molecular biology (Clifton, N.J.) 2013

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Transfection of mammalian cells has long been an extremely powerful approach for the study of the effects of specific gene expression on cell function. Until recently, however, this approach has been unavailable for the study of gene function in adult cardiac myocytes. Here, an adaptation of the biolistic method to the transfection of adult cardiac myocytes is described. DNA is precipitated onto gold particles in the absence of PVP and the particles are biolistically delivered to freshly isolated adult rat cardiomyocytes via a Bio-Rad Helios System gene gun. The myocytes are cultured in the absence of bovine serum albumin and expression of the introduced genes, in phenotypically intact myocytes, is robust within 12-24 h.

Citation

David F Steele, Ying Dou, David Fedida. Biolistic transfection of freshly isolated adult ventricular myocytes. Methods in molecular biology (Clifton, N.J.). 2013;940:145-55