Generation and Characterization of Patient-Specific iPSC Model for Cardiovascular Disease

Methods Mol Biol. 2016;1353:191-213. doi: 10.1007/7651_2015_273.

Abstract

Advances in differentiation of cardiomyocytes from human induced pluripotent stem cell (hiPSC) were emerged as a tool for modeling of cardiovascular disease that recapitulates the phenotype for the purpose of drug screening, biomarker discovery, and testing of single-nucleotide polymorphism (SNP) as a modifier for disease stratification. Here, we describe the (1) retroviral reprogramming strategies in the generation of human iPSC, (2) methodology in characterization of iPSC in order to identify the stem cell clones with the best quality, and (3) protocol of cardiac differentiation by modulation of Wnt signaling and β-catenin pathway.

Keywords: Cardiac differentiation; Cardiovascular disease modeling; Human induced pluripotent stem cell (hiPSC); Retroviral reprogramming; Stem cell characterization.

MeSH terms

  • Amides / pharmacology
  • Animals
  • Biomarkers / metabolism
  • Cell Culture Techniques / methods*
  • Cell Differentiation
  • Cellular Reprogramming*
  • Collagen / chemistry
  • Drug Combinations
  • Embryo, Mammalian
  • Embryoid Bodies / cytology*
  • Embryoid Bodies / drug effects
  • Embryoid Bodies / metabolism
  • Enzyme Inhibitors / pharmacology
  • Feeder Cells / cytology
  • Fibroblasts / cytology
  • Gene Expression
  • Genetic Vectors
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Laminin / chemistry
  • Mice
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism
  • Primary Cell Culture
  • Proteoglycans / chemistry
  • Pyridines / pharmacology
  • Pyrimidines / pharmacology
  • Retroviridae / genetics
  • Teratoma / genetics
  • Teratoma / metabolism
  • Teratoma / pathology
  • Wnt Signaling Pathway
  • beta Catenin / genetics
  • beta Catenin / metabolism

Substances

  • Amides
  • Biomarkers
  • CTNNB1 protein, human
  • Chir 99021
  • Drug Combinations
  • Enzyme Inhibitors
  • Intercellular Signaling Peptides and Proteins
  • Laminin
  • Proteoglycans
  • Pyridines
  • Pyrimidines
  • beta Catenin
  • matrigel
  • Y 27632
  • Collagen