title: Isolation and characterization of a Sca-1+/CD31- progenitor cell lineage derived from mouse heart tissue.
abstract: BACKGROUND
  Myocardial infarction remains the leading cause of mortality in developed countries despite recent advances in its prevention and treatment. Regenerative therapies based on resident cardiac progenitor cells (CPCs) are a promising alternative to conventional treatments. However, CPCs resident in the heart are quite rare. It is unclear how these CPCs can be isolated and cultured efficiently and what the effects of long-term culture in vitro are on their 'stemness' and differentiation potential, but this is critical knowledge for CPCs' clinical application.
  
  
  RESULTS
  Here, we isolated stem cell antigen-1 positive cells from postnatal mouse heart by magnetic active cell sorting using an iron-labeled anti-mouse Sca-1 antibody, and cultured them long-term in vitro. We tested stemness marker expression and the proliferation ability of long-term cultured Sca-1+ cells at early, middle and late passages. Furthermore, we determined the differentiation potential of these three passages into cardiac cell lineages (cardiomyocytes, smooth muscle and endothelial cells) after induction in vitro. The expression of myocardial, smooth muscle and endothelial cell-specific genes and surface markers were analyzed by RT-PCR and IF staining. We also investigated the oncogenicity of the three passages by subcutaneously injecting cells in nude mice. Overall, heart-derived Sca-1+ cells showed CPC characteristics: long-term propagation ability in vitro, non-tumorigenic in vivo, persistent expression of stemness and cardiac-specific markers, and multipotent differentiation into cardiac cell lineages.
  
  
  CONCLUSIONS
  Our research may bring new insights to myocardium regeneration, for which even a small number of biopsy-derived CPCs could be enriched and propagated long term in vitro to obtain sufficient seed cells for cell injection or cardiac tissue engineering.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.forename: Bei
authors.initials: B
authors.lastname: Feng
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.forename: Xiaomin
authors.initials: X
authors.lastname: He
authors.forename: Renjie
authors.initials: R
authors.lastname: Hu
authors.forename: Meng
authors.initials: M
authors.lastname: Yin
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Pediatric Cardiothoracic Surgery, Shanghai Children's Medical Center, School of Medicine, Shanghai Jiao Tong University, 1678 Dong Fang Road, Shanghai 200127, China. fuweizhulu@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Fu
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Xu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000950:Antigens, Ly / Q000276:immunology / Q000378:metabolism*; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D007501:Iron / Q000737:chemistry; D008297:Male; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008813:Mice, Inbred ICR; D008819:Mice, Nude; D009206:Myocardium / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D059467:Transcriptome
chemical_list: D000906:Antibodies; D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D007501:Iron
references.citation: Am J Cardiol. 1991 Nov 18;68(14):1D-6D
references.pmid: 1836091
references.citation: Circ Res. 2004 Oct 29;95(9):911-21
references.pmid: 15472116
references.citation: Circulation. 2008 Sep 30;118(14 Suppl):S145-52
references.pmid: 18824746
references.citation: Circulation. 2007 Feb 20;115(7):896-908
references.pmid: 17283259
references.citation: Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8626-31
references.pmid: 11447262
references.citation: Methods Mol Biol. 2013;1036:63-74
references.pmid: 23807786
references.citation: Circulation. 2010 Feb 23;121(7):948-54
references.pmid: 20177011
references.citation: Curr Opin Cardiol. 2000 May;15(3):178-82
references.pmid: 10952425
references.citation: J Cell Sci. 2007 May 15;120(Pt 10):1791-800
references.pmid: 17502484
references.citation: Physiol Rev. 2005 Oct;85(4):1373-416
references.pmid: 16183916
references.citation: Science. 2009 Oct 16;326(5951):426-9
references.pmid: 19833966
references.citation: J Cell Biol. 2007 Jan 29;176(3):329-41
references.pmid: 17261849
references.citation: J Clin Invest. 2003 May;111(10):1497-504
references.pmid: 12750399
references.citation: Circ Res. 2005 Jul 8;97(1):52-61
references.pmid: 15947249
references.citation: Science. 1994 Apr 1;264(5155):98-101
references.pmid: 8140423
references.citation: Blood. 2009 Jul 9;114(2):268-78
references.pmid: 19420357
references.citation: Nature. 2009 Jul 2;460(7251):113-7
references.pmid: 19571884
references.citation: J Control Release. 2013 Aug 10;169(3):336-40
references.pmid: 23500057
references.citation: Dev Cell. 2003 Dec;5(6):877-89
references.pmid: 14667410
references.citation: Nature. 2005 Feb 10;433(7026):647-53
references.pmid: 15703750
references.citation: J Immunol. 1998 Apr 15;160(8):3861-8
references.pmid: 9558091
references.citation: J Biol Chem. 2004 Mar 19;279(12):11384-91
references.pmid: 14702342
references.citation: Cell. 2006 Dec 15;127(6):1137-50
references.pmid: 17123591
references.citation: Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12313-8
references.pmid: 14530411
references.citation: Cell. 2006 Dec 15;127(6):1151-65
references.pmid: 17123592
references.citation: Nat Protoc. 2009;4(2):232-43
references.pmid: 19197267
references.citation: Nature. 2011 Jun 08;474(7353):640-4
references.pmid: 21654746
references.citation: PLoS One. 2011;6(9):e25265
references.pmid: 21980409
references.citation: Stem Cells. 2009 Jul;27(7):1571-81
references.pmid: 19544463
references.citation: Cell. 2003 Sep 19;114(6):763-76
references.pmid: 14505575