title: Cardiomyocyte differentiation from mouse embryonic stem cells using a simple and defined protocol.
abstract: BACKGROUND
  Embryonic stem (ES) cells are pluripotent cells with the ability to differentiate to any cell type of the resident organism. In recent years, significant advances have been made in using these cells to obtain large numbers of cardiomyocyte (CM)-like cells for scientific research and clinical application. A vast number of protocols have emerged describing differentiation methods without the use of animal serum or extracts restrictive for use in a human clinical setting. These techniques follow a complicated procedure, which although successful, show a relatively varied yield among cell batches.
  
  
  RESULTS
  We have designed a three-step differentiation protocol using defined reagents and a monolayer culture without feeder cells, avoiding embryoid body formation and multiple trypsin treatment, in which beating foci appeared as early as day 6 in in vitro differentiating conditions. Our results show a high yield of CM reaching approximately 60% of the differentiated cells after 13 days in vitro.
  
  
  CONCLUSIONS
  We provide a fast, simple, reliable and reproducible protocol for inducing murine ES cells toward a CM-like phenotype comparable to available high-yield protocols, without the use of intermediate trypsinization/passage steps.
authors.affiliation: Translational Medicine and Therapeutics, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, United Kingdom.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Kokkinopoulos
authors.affiliation: Translational Medicine and Therapeutics, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, United Kingdom.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Ishida
authors.affiliation: Translational Medicine and Therapeutics, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, United Kingdom.
authors.forename: Rie
authors.initials: R
authors.lastname: Saba
authors.affiliation: Translational Medicine and Therapeutics, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, United Kingdom.
authors.forename: Steven
authors.initials: S
authors.lastname: Coppen
authors.affiliation: Translational Medicine and Therapeutics, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, United Kingdom.
authors.forename: Ken
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Translational Medicine and Therapeutics, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, United Kingdom.
authors.forename: Kenta
authors.initials: K
authors.lastname: Yashiro
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology*
keywords: cardiomyocyte differentiation; feeder-free; monolayer culture; mouse embryonic stem cell; serum-free