TRANSFER OF MACROMOLECULES INTO LIVING ADULT CARDIOMYOCYTES BY MICROINJECTION

Among techniques commonly used to deliver bioactive molecules into liv ing cells, microinjection is a very efficient method. Microinjection h as been used extensively for gene transfer into different cell types. We applied the microinjection technique to the adult rat ventricular c ardiac muscle cells (AVC) in primary culture and optimized microinject ion parameters and the appropriate cell culture conditions. We also op timized the use of particular agents (i.e. 2,3-butanedione monoxime, v erapamil) for the prevention of the cell damage caused by the micropun cture. We obtained the expression of a CMV-beta-galactosidase reporter gene in up to 20% of the injected cells with efficient maintenance of long term cell viability. Under our experimental conditions direct mi croinjection is a very advantageous technique to transfer macromolecul es into living adult cardiac muscle cells and a powerful system to stu dy and manipulate the biochemistry and molecular biology of the cardia c myocyte.