title: The long-term differentiation of embryonic stem cells into cardiomyocytes: an indirect co-culture model.
abstract: BACKGROUND
  Embryonic Stem Cells (ESCs) can differentiate into cardiomyocytes (CMs) in vitro but the differentiation level from ESCs is low. Here we describe a simple co-culture model by commercially available Millicell™ hanging cell culture inserts to control the long-term differentiation of ESCs into CMs.
  
  
  RESULTS
  Mouse ESCs were cultured in hanging drops to form embryoid bodies (EBs) and treated with 0.1 mmol/L ascorbic acid to induce the differentiation of ESCs into CMs. In the indirect co-culture system, EBs were co-cultured with epidermal keratinocytes (EKs) or neonatal CMs (NCMs) by the hanging cell culture inserts (PET membranes with 1 µm pores). The molecular expressions and functional properties of ESC-derived CMs in prolonged culture course were evaluated. During time course of ESC differentiation, the percentages of EBs with contracting areas in NCMs co-culture were significantly higher than that without co-culture or in EKs co-culture. The functional maintenance of ESC-derived CMs were more prominent in NCMs co-culture model.
  
  
  CONCLUSIONS
  These results indicate that NCMs co-culture promote ESC differentiation and has a further effect on cell growth and differentiation. We assume that the improvement of the differentiating efficiency of ESCs into CMs in the co-culture system do not result from the effect of co-culture directly on cell differentiation, but rather by signaling effects that influence the cells in proliferation and long-term function maintenance.
authors: Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi'an, China.
authors: Dong-Bo
authors: DB
authors: Ou
authors: Di
authors: D
authors: Zeng
authors: Yan
authors: Y
authors: Jin
authors: Xiong-Tao
authors: XT
authors: Liu
authors: Ji-Wei
authors: JW
authors: Teng
authors: Wan-Gang
authors: WG
authors: Guo
authors: Hong-Tao
authors: HT
authors: Wang
authors: Fei-Fei
authors: FF
authors: Su
authors: Yong
authors: Y
authors: He
authors: Qiang-Sun
authors: QS
authors: Zheng
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D018920:Coculture Techniques / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*
chemical_list: D001205:Ascorbic Acid
references: Cardiovasc Res. 2010 Nov 1;88(2):314-23
references: 20547733
references: Br J Haematol. 1995 Jul;90(3):518-25
references: 7646988
references: Cell. 2007 Jan 26;128(2):325-39
references: 17254970
references: Differentiation. 1991 Dec;48(3):173-82
references: 1725163
references: Genes Dev. 1997 Apr 15;11(8):1061-72
references: 9136933
references: Circ Res. 2009 Feb 27;104(4):e30-41
references: 19213953
references: J Physiol. 2007 Dec 1;585(Pt 2):565-78
references: 17932143
references: Cardiovasc Res. 2009 Nov 1;84(2):263-72
references: 19564151
references: PLoS One. 2010 Jun 15;5(6):e11134
references: 20559569
references: Anat Rec. 2002 Feb 1;266(2):93-102
references: 11788942
references: Pflugers Arch. 1999 Apr;437(5):669-79
references: 10087143
references: Ann Hematol. 2005 Oct;84(11):715-21
references: 16096830
references: Int J Cancer. 1995 Jan 3;60(1):93-9
references: 7529213
references: FASEB J. 2004 Nov;18(14):1710-2
references: 15345687
references: Differentiation. 2008 Apr;76(4):357-70
references: 18021257
references: FASEB J. 2002 Oct;16(12):1558-66
references: 12374778
references: Differentiation. 2006 Apr;74(4):149-59
references: 16683985
references: Exp Cell Res. 2007 Mar 10;313(5):921-30
references: 17258200
references: Circulation. 2003 Apr 15;107(14):1912-6
references: 12668514
references: Science. 1998 Nov 6;282(5391):1145-7
references: 9804556
references: J Cell Biochem. 2011 Dec;112(12):3555-62
references: 21780160
references: FASEB J. 1997 Sep;11(11):905-15
references: 9285489
references: Circ Res. 1999 Feb 5;84(2):136-45
references: 9933244
references: Bioessays. 2009 Feb;31(2):246-52
references: 19204979
references: Tissue Eng Part A. 2010 Apr;16(4):1349-61
references: 19908964
references: PLoS One. 2009;4(4):e5046
references: 19352491
references: Circ Res. 1994 Aug;75(2):233-44
references: 8033337
references: PLoS One. 2010 Jul 12;5(7):e11536
references: 20634944
references: Development. 1990 Dec;110(4):1341-8
references: 2129226
references: Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11307-12
references: 11027332
references: Stem Cells. 2007 Apr;25(4):929-38
references: 17185609
references: Int J Cardiol. 2007 Apr 12;117(1):80-9
references: 17034884
references: Circulation. 2003 Jun 3;107(21):2733-40
references: 12742992
references: Stem Cells. 2007 Mar;25(3):553-61
references: 17332514