title: Ascorbic Acid-Induced Cardiac Differentiation of Murine Pluripotent Stem Cells: Transcriptional Profiling and Effect of a Small Molecule Synergist of Wnt/β-Catenin Signaling Pathway.
abstract: BACKGROUND
  Reproducible and efficient differentiation of pluripotent stem cells (PSCs) to cardiomyocytes (CMs) is essential for their use in regenerative medicine, drug testing and disease modeling. The aim of this study was to evaluate the effect of some previously reported cardiogenic substances on cardiac differentiation of mouse PSCs.
  
  
  METHODS
  Differentiation was performed by embryoid body (EB)-based method using three different murine PSC lines. The differentiation efficiency was monitored by RT-qPCR, immunocytochemistry and flow cytometry, and the effect mechanistically evaluated by transcriptome analysis of treated EBs.
  
  
  RESULTS
  Among the five tested compounds (ascorbic acid, dorsomorphin, cyclic adenosine 3',5'-monophosphate, cardiogenol C, cyclosporin A) only ascorbic acid (AA) exerted a strong and reproducible cardiogenic effect in CGR8 cells which was less consistent in other two PSC lines. AA induced only minor changes in transcriptome of CGR8 cells after administration during the initial two days of differentiation. Cardiospecific genes and transcripts involved in angiogenesis, erythropoiesis and hematopoiesis were up-regulated on day 5 but not on days 2 or 3 of differentiation. The cardiac differentiation efficiency was improved when QS11, a small-molecule synergist of Wnt/β-catenin signaling pathway, was added to cultures after AA-treatment.
  
  
  CONCLUSIONS
  This study demonstrates that only minor transcriptional changes are sufficient for enhancement of cardiogenesis of murine PSCs by AA and that AA and QS11 exhibit synergistic effects and enhance the efficiency of CM differentiation of murine PSCs.
authors.affiliation: Center for Physiology and Pathophysiology, Institute for Neurophysiology, Medical Faculty, University of Cologne, Cologne, Germany.
authors.forename: Dina
authors.initials: D
authors.lastname: Ivanyuk
authors.forename: Galina
authors.initials: G
authors.lastname: Budash
authors.forename: Yunjie
authors.initials: Y
authors.lastname: Zheng
authors.forename: John Antony
authors.initials: JA
authors.lastname: Gaspar
authors.forename: Umesh
authors.initials: U
authors.lastname: Chaudhari
authors.forename: Azra
authors.initials: A
authors.lastname: Fatima
authors.forename: Soghra
authors.initials: S
authors.lastname: Bahmanpour
authors.forename: Vladislav K
authors.initials: VK
authors.lastname: Grin
authors.forename: Andrey G
authors.initials: AG
authors.lastname: Popandopulo
authors.forename: Agapios
authors.initials: A
authors.lastname: Sachinidis
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hescheler
authors.forename: Tomo
authors.initials: T
authors.lastname: Šarić
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D020869:Gene Expression Profiling; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011687:Purines / Q000494:pharmacology; D060449:Wnt Signaling Pathway / Q000187:drug effects*; D051176:beta Catenin / Q000378:metabolism
chemical_list: D000975:Antioxidants; D011687:Purines; C520339:QS11 compound; D051176:beta Catenin; D001205:Ascorbic Acid