title: Cardiomyocyte marker expression in mouse embryonic fibroblasts by cell-free cardiomyocyte extract and epigenetic manipulation.
abstract: BACKGROUND
  The regenerative capacity of the mammalian heart is quite limited. Recent reports have focused on reprogramming mesenchymal stem cells into cardiomyocytes. We investigated whether fibroblasts could transdifferentiate into myocardium.
  
  
  METHODS
  Mouse embryonic fibroblasts were treated with Trichostatin A (TSA) and 5-Aza-2-Deoxycytidine (5-aza-dC). The treated cells were permeabilized with streptolysin O and exposed to the mouse cardiomyocyte extract and cultured for 1, 10, and 21 days. Cardiomyocyte markers were detected by immunohistochemistry. Alkaline phosphatase activity and OCT4 were also detected in cells treated by chromatin-modifying agents.
  
  
  RESULTS
  The cells exposed to a combination of 5-aza-dC and TSA and permeabilized in the presence of the cardiomyocyte extract showed morphological changes. The cells were unable to express cardiomyocyte markers after 24 h. Immunocytochemical assays showed a notable degree of myosin heavy chain and α-actinin expressions after 10 days. The expression of the natriuretic factor and troponin T occurred after 21 days in these cells. The cells exposed to chromatin-modifying agents also expressed cardiomyocyte markers; however, the proportion of reprogrammed cells was clearly smaller than that in the cultures exposed to 5-aza-dC , TSA, and extract.
  
  
  CONCLUSIONS
  It seems that the fibroblasts were able to eliminate the previous epigenetic markers and form new ones according to the factors existing in the extract. Since no beating was observed, at least up to 21 days, the cells may need an appropriate extracellular matrix for their function.
authors.affiliation: Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; ; Department of Tissue Engineering, School of Advance Sciences and Technology, Shiraz University of Medical Sciences, Shiraz, Iran;
authors.forename: Tahereh
authors.initials: T
authors.lastname: Talaei-Khozani
authors.affiliation: Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran;
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Heidari
authors.affiliation: Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran;
authors.forename: Tahereh
authors.initials: T
authors.lastname: Esmaeilpour
authors.affiliation: Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran;
authors.forename: Zahra
authors.initials: Z
authors.lastname: Vojdani
authors.affiliation: Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Zohrah
authors.initials: Z
authors.lastname: Mostafavi-Pour
authors.affiliation: Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran;
authors.forename: Leili
authors.initials: L
authors.lastname: Rohani
keywords: Cardiomyocytes; Cell transdifferentiation; Fibroblast; Histone deacetylase inhibitors
references.citation: J Thorac Cardiovasc Surg. 2003 Jul;126(1):124-32
references.pmid: 12878947
references.citation: Hum Cell. 2011 Mar;24(1):35-42
references.pmid: 21547694
references.citation: Circulation. 2003 Feb 25;107(7):1024-32
references.pmid: 12600917
references.citation: Mol Biol Rep. 2010 Mar;37(3):1197-202
references.pmid: 19263240
references.citation: J Pharmacol Exp Ther. 2012 Apr;341(1):285-93
references.pmid: 22271820
references.citation: J Cardiovasc Transl Res. 2013 Feb;6(1):37-45
references.pmid: 23054660
references.citation: Development. 2012 Dec 1;139(23):4330-40
references.pmid: 23132243
references.citation: Nat Biotechnol. 2002 May;20(5):460-6
references.pmid: 11981558
references.citation: Front Biosci (Schol Ed). 2012 Jun 01;4:1413-23
references.pmid: 22652883
references.citation: Nat Biotechnol. 2008 Nov;26(11):1269-75
references.pmid: 18849973
references.citation: EMBO Rep. 2002 Apr;3(4):384-9
references.pmid: 11897658
references.citation: Differentiation. 2010 Feb;79(2):93-101
references.pmid: 19926393
references.citation: Mol Biol Cell. 2005 Dec;16(12):5719-35
references.pmid: 16195347
references.citation: Cell Biol Int. 2009 Nov;33(11):1184-93
references.pmid: 19729070
references.citation: Cytotherapy. 2010 May;12(3):332-7
references.pmid: 20230311
references.citation: Nature. 2008 Jul 3;454(7200):49-55
references.pmid: 18509334
references.citation: Scand J Clin Lab Invest. 2008;68(6):464-72
references.pmid: 18609095
references.citation: Circulation. 2002 Jan 1;105(1):93-8
references.pmid: 11772882
references.citation: Sci China Life Sci. 2010 May;53(5):581-9
references.pmid: 20596941
references.citation: Zygote. 2011 Aug;19(3):237-43
references.pmid: 20569515
references.citation: Cancer Cell. 2008 Sep 9;14(3):193-4
references.pmid: 18772107
references.citation: Biochem Soc Trans. 2010 Apr;38(2):370-3
references.pmid: 20298185
references.citation: Biochem Biophys Res Commun. 2004 Apr 9;316(3):834-41
references.pmid: 15033476
references.citation: Cloning Stem Cells. 2008 Sep;10(3):371-9
references.pmid: 18419249
references.citation: Biochem Biophys Res Commun. 2004 Feb 6;314(2):420-7
references.pmid: 14733922
references.citation: Nature. 2009 Jun 4;459(7247):708-11
references.pmid: 19396158
references.citation: Exp Biol Med (Maywood). 2004 Jul;229(7):623-31
references.pmid: 15229356
references.citation: Dev Biol. 2003 Jun 15;258(2):307-18
references.pmid: 12798290
references.citation: Cell Mol Life Sci. 2007 May;64(9):1043-62
references.pmid: 17347798
references.citation: Theriogenology. 2009 Nov;72(8):1097-110
references.pmid: 19765811
references.citation: Iran J Reprod Med. 2012 May;10(3):193-200
references.pmid: 25242993
references.citation: Mol Cells. 2007 Dec 31;24(3):441-4
references.pmid: 18182862
references.citation: J Mol Cell Cardiol. 1979 May;11(5):423-33
references.pmid: 448740
references.citation: Curr Med Chem. 2004 Jun;11(12):1629-43
references.pmid: 15180568
references.citation: Stem Cells. 2008 Nov;26(11):2884-92
references.pmid: 18687994
references.citation: Results Probl Cell Differ. 2012;55:291-302
references.pmid: 22918813
references.citation: Exp Cell Res. 2008 Aug 15;314(14):2634-42
references.pmid: 18571647
references.citation: Cancer Lett. 2007 May 18;250(1):53-62
references.pmid: 17189669