title: Derivation of cardiac progenitor cells from embryonic stem cells.
abstract: Cardiac progenitor cells (CPCs) have the capacity to differentiate into cardiomyocytes, smooth muscle cells (SMC), and endothelial cells and hold great promise in cell therapy against heart disease. Among various methods to isolate CPCs, differentiation of embryonic stem cell (ESC) into CPCs attracts great attention in the field since ESCs can provide unlimited cell source. As a result, numerous strategies have been developed to derive CPCs from ESCs. In this protocol, differentiation and purification of embryonic CPCs from both mouse and human ESCs is described. Due to the difficulty of using cell surface markers to isolate embryonic CPCs, ESCs are engineered with fluorescent reporters activated by CPC-specific cre recombinase expression. Thus, CPCs can be enriched by fluorescence-activated cell sorting (FACS). This protocol illustrates procedures to form embryoid bodies (EBs) from ESCs for CPC specification and enrichment. The isolated CPCs can be subsequently cultured for cardiac lineage differentiation and other biological assays. This protocol is optimized for robust and efficient derivation of CPCs from both mouse and human ESCs.
authors: Cardiac Surgery, University of Michigan.
authors: Ieng Lam
authors: IL
authors: Lei
authors: Leon H Charney Division of Cardiology, New York University School of Medicine.
authors: Lei
authors: L
authors: Bu
authors: Cardiac Surgery, University of Michigan; zhongw@med.umich.edu.
authors: Zhong
authors: Z
authors: Wang
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology*; D042783:Endothelial Cells / Q000166:cytology; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology
references: Science. 2013 Feb 15;339(6121):823-6
references: 23287722
references: Nature. 2008 May 15;453(7193):322-9
references: 18480813
references: Cell Stem Cell. 2013 Jun 6;12(6):689-98
references: 23746978
references: Trends Biotechnol. 2013 Jul;31(7):397-405
references: 23664777
references: Proc Natl Acad Sci U S A. 2013 Jul 30;110(31):12667-72
references: 23861494
references: Nat Cell Biol. 2013 Sep;15(9):1098-106
references: 23974038
references: Cell Stem Cell. 2008 Jul 3;3(1):69-84
references: 18593560
references: Differentiation. 2008 Nov;76(9):958-70
references: 18557764
references: Nature. 2009 Jul 2;460(7251):113-7
references: 19571884
references: Circ Res. 2010 Dec 10;107(12):1414-27
references: 21148448
references: Cell Stem Cell. 2011 Feb 4;8(2):228-40
references: 21295278
references: PLoS One. 2011;6(4):e18293
references: 21494607
references: Proc Natl Acad Sci U S A. 2012 Jul 3;109(27):E1848-57
references: 22645348
references: J Biol Chem. 2012 Jul 13;287(29):24255-62
references: 22621927
references: Cell. 2012 Sep 28;151(1):206-20
references: 22981692
references: Nat Rev Mol Cell Biol. 2013 Jan;14(1):49-55
references: 23169466
references: Science. 2013 Feb 15;339(6121):819-23
references: 23287718
references: J Cell Biochem. 2013 Nov;114(11):2437-45
references: 23606236
references: Cell. 2013 Sep 12;154(6):1370-9
references: 23992847
references: Stem Cell Res. 2013 Nov;11(3):1335-47
references: 24095945
references: Circ Res. 2002 Aug 9;91(3):189-201
references: 12169644
references: Circulation. 2003 Apr 15;107(14):1912-6
references: 12668514
references: Proc Natl Acad Sci U S A. 2005 Sep 13;102(37):13170-5
references: 16141334
references: Dev Biol. 2005 Nov 1;287(1):134-45
references: 16188249
references: Cell. 2006 Dec 15;127(6):1137-50
references: 17123591
references: Nat Biotechnol. 2007 Sep;25(9):1015-24
references: 17721512
references: Cell Stem Cell. 2013 Apr 4;12(4):393-4
references: 23561441