9 - Primary Cultures of Cardiac Myocytes as In Vitro Models for Pharmacological and Toxicological Assessments

Abstract

Primary cultures of cardiac myocytes are attractive models that can minimize the use of animals and reduce the cost of developing cardioactive compounds. It describes methodologies for the preparation of primary cultures of neonatal rat cardiac myocytes, adult rabbit cardiac myocytes, and several end-point assays that can be used to assess pharmacological and toxicological responses of cardioactive compounds. These models have a biological basis that links them to the in vivo biological response being modeled. The chapter reviews different assays that could be used to assess pharmacological and toxicological responses of cardiac myocytes in culture. These assays are essentially divided into two categories: those that measure alterations of physiological functions and those that measure cell death. Cell death is an abrupt all-or-nothing phenomenon that occurs as the consequence of abrupt breakdown of the plasma membrane. Once the dose and time dependency of cell death have been established, functional assays can proceed to identify the molecular and cellular mechanisms of action. In addition, this review describes new technologies in fluorescence microscopy that provide powerful tools to investigate mechanisms of actions.