Adenovirus-mediated myofilament gene transfer into adult cardiac myocytes.
作者： Margaret V. Westfall , Elizabeth M. Rust , Faris Albayya , Joseph M. Metzger
DOI: 10.1016/S0091-679X(08)60385-4
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摘要: Publisher Summary This chapter provides methods for the isolation of Ca2+-tolerant, rod-shaped adult cardiac myocytes suitable adenovirus-mediated myofilament gene transfer; construction recombinant adenovirus vectors; and detection ectopic protein expression incorporation in contractile apparatus. Recombinant vectors can be used to accomplish highly efficient transfer, expression, into ventricular primary culture. approach is made possible by a culture system that allows maintenance both cellular subcellular differentiated state are constructed homologous recombination shuttle vector containing desired cDNA with plasmid full-length DNA human embryonic kidney (HEK 293) cell line. The then plaque purified high-titer virus grown from an individual subsequent infection isolated myocytes.
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