Maass Alexander H, Buvoli Massimo
Department of Medicine, University of Wuerzburg, Germany.
Methods Mol Biol. 2007;366:321-30. doi: 10.1007/978-1-59745-030-0_18.
Neonatal rat ventricular myocytes (NRVMs) cultured in vitro have been used as a model system for easily recreating and studying several cardiac molecular conditions, such as hypertrophy, oxygen deprivation, and gene expression. However, low efficiency of gene transfer has often represented one of the major limitations of this technique. In this chapter we describe in detail how to isolate NRVMs from neonatal rat heart and the optimal conditions for their long-term culture. Different cardiomyocyte transfection methodologies, based on viral or viral/chemical delivery carriers, are also discussed.
体外培养的新生大鼠心室肌细胞(NRVMs)已被用作一种模型系统,用于轻松再现和研究多种心脏分子状况,如肥大、缺氧和基因表达。然而,基因转移效率低常常是该技术的主要局限之一。在本章中,我们将详细描述如何从新生大鼠心脏中分离NRVMs以及长期培养它们的最佳条件。还将讨论基于病毒或病毒/化学递送载体的不同心肌细胞转染方法。