title: Simple Monolayer Differentiation of Murine Cardiomyocytes via Nutrient Deprivation-Mediated Activation of β-Catenin.
abstract: Methods to generate murine cardiomyocytes from pluripotent stem cells (PSCs) in vitro are resource and time intensive. All current protocols require exogenously provided soluble factors and almost all utilize embryoid body formation to modulate pathways associated with mesoderm specification and cardiomyocyte differentiation. Here, we developed a simple protocol without EBs and without exogenous soluble factors that enabled cardiomyocyte differentiation of a murine induced PSC line based on controlled nutrient deprivation in 2D monolayer cultures. We showed that this protocol reproducibly imposed metabolic stress and consequently modulated active β-catenin levels to yield functional cardiomyocytes. The yield of cardiomyocytes and calcium handling kinetics were comparable to existing approaches. However, this approach did not produce consistent results between murine PSC lines suggesting signaling pathways linking nutrient deprivation to β-catenin activation are not universally conserved and may be a remnant of the parent population from which the induced PSCs were derived.
authors.affiliation: Department of Biomedical Engineering, University of Minnesota - Twin Cities, 312 Church St. SE, Minneapolis, MN, 55455, USA.|Department of Biochemical Engineering, University of Applied Sciences Technikum Wien, Höchstädtplatz 5, 1200, Vienna, Austria.
authors.forename: Pablo
authors.initials: P
authors.lastname: Hofbauer
authors.affiliation: Department of Biomedical Engineering, University of Minnesota - Twin Cities, 312 Church St. SE, Minneapolis, MN, 55455, USA.|Stem Cell Institute, University of Minnesota -Twin Cities, Minneapolis, 55455, MN, USA.
authors.forename: Jangwook P
authors.initials: JP
authors.lastname: Jung
authors.affiliation: Department of Biomedical Engineering, University of Minnesota - Twin Cities, 312 Church St. SE, Minneapolis, MN, 55455, USA.
authors.forename: Tanner J
authors.initials: TJ
authors.lastname: McArdle
authors.affiliation: Department of Biomedical Engineering, University of Minnesota - Twin Cities, 312 Church St. SE, Minneapolis, MN, 55455, USA. ogle@umn.edu.|Stem Cell Institute, University of Minnesota -Twin Cities, Minneapolis, 55455, MN, USA. ogle@umn.edu.|Masonic Cancer Center, University of Minnesota - Twin Cities, 312 Church St. SE, Minneapolis, MN, 55455, USA. ogle@umn.edu.|Lillehei Heart Institute, University of Minnesota - Twin Cities, 312 Church St. SE, Minneapolis, MN, 55455, USA. ogle@umn.edu.|Institute for Engineering in Medicine, University of Minnesota - Twin Cities, 312 Church St. SE, Minneapolis, MN, 55455, USA. ogle@umn.edu.
authors.forename: Brenda M
authors.initials: BM
authors.lastname: Ogle
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D019344:Lactic Acid / Q000378:metabolism; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D019289:Pyruvic Acid / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors; D060449:Wnt Signaling Pathway / Q000235:genetics; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism*
chemical_list: D051176:beta Catenin; D019344:Lactic Acid; D019289:Pyruvic Acid; D005947:Glucose; D002118:Calcium
keywords: Metabolism; Murine PSCs; Nutrient deprivation; Simple cardiomyocyte differentiation; Wnt-signaling