title: Mouse embryonic stem cell-derived cardiac myocytes in a cell culture dish.
abstract: Embryonic stem (ES) cells are pluripotent stem cells capable of self-renewal and have broad differentiation potential yielding cell types from all three germ layers. In the absence of differentiation inhibitory factors, when cultured in suspension, ES cells spontaneously differentiate and form three-dimensional cell aggregates termed embryoid bodies (EBs). Although various methods exist for the generation of EBs, the hanging drop method offers reproducibility and homogeneity from a predetermined number of ES cells. Herein, we describe the in vitro differentiation of mouse embryonic stem cells into cardiac myocytes using the hanging drop method and immunocytochemistry to identify cardiomyogenic differentiation. In brief, ES cells, placed in droplets on the lid of culture dishes following a 2-day incubation, yield embryoid bodies, which are resuspended and plated. 1-2 weeks following plating of the EBs, spontaneous beating areas can be observed and staining for specific cardiac markers can be achieved.
authors.affiliation: Biomolecular Science Center, Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4110 Libra Drive, Building 20 Room 320A, Orlando, FL, 32817, USA.
authors.forename: Carley
authors.initials: C
authors.lastname: Glass
authors.forename: Reetu
authors.initials: R
authors.lastname: Singla
authors.forename: Anshu
authors.initials: A
authors.lastname: Arora
authors.forename: Dinender K
authors.initials: DK
authors.lastname: Singla
mesh_terms: D000818:Animals; D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D058732:Embryoid Bodies; D007150:Immunohistochemistry; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism
chemical_list: D015415:Biomarkers