Calcium Imaging in Pluripotent Stem Cell-Derived Cardiac Myocytes

Methods Mol Biol. 2016:1353:131-46. doi: 10.1007/7651_2015_267.

Abstract

The possibility to generate cardiomyocytes (CMs) from disease-specific induced pluripotent stem cells (iPSCs) is a powerful tool for the investigation of various cardiac diseases in vitro. The pathological course of various cardiac conditions, causatively heterogeneous, often converges into disturbed cellular Ca(2+) cycling. The gigantic Ca(2+) channel of the intracellular Ca(2+) store of CMs, the ryanodine receptor type 2 (RyR2), controls Ca(2+) release and therefore plays a crucial role in Ca(2+) cycling of CMs. In the present protocol we describe ways to measure and analyze global as well as local cellular Ca(2+) release events in CMs derived from a patient carrying a CPVT-causing RyR2 mutation.

Keywords: Calcium transients; Cardiomyocytes; Confocal microscopy; Sparks; Tachycardia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aniline Compounds
  • Animals
  • Calcium / metabolism
  • Calcium Signaling*
  • Cell Culture Techniques / methods*
  • Cell Differentiation
  • Cellular Reprogramming*
  • Fibroblasts / cytology
  • Fibroblasts / metabolism*
  • Gene Expression
  • Humans
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism*
  • Induced Pluripotent Stem Cells / ultrastructure
  • Isoproterenol / pharmacology
  • Mice
  • Molecular Imaging / methods
  • Mutation
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism*
  • Myocytes, Cardiac / ultrastructure
  • Primary Cell Culture
  • Ryanodine / pharmacology
  • Ryanodine Receptor Calcium Release Channel / genetics
  • Ryanodine Receptor Calcium Release Channel / metabolism
  • Tachycardia / genetics
  • Tachycardia / metabolism
  • Tachycardia / pathology
  • Xanthenes

Substances

  • Aniline Compounds
  • Fluo 4
  • Ryanodine Receptor Calcium Release Channel
  • Xanthenes
  • Ryanodine
  • Isoproterenol
  • Calcium