title: Preparation of cardiac extracellular matrix from an intact porcine heart.
abstract: Whole organ engineering would benefit from a three-dimensional scaffold produced from intact organ-specific extracellular matrix (ECM). The microenvironment and architecture provided by such a scaffold would likely support site-appropriate cell differentiation and spatial organization. The methods to produce such scaffolds from intact organs require customized decellularization protocols. In the present study, intact adult porcine hearts were successfully decellularized in less than 10 h using pulsatile retrograde aortic perfusion. Serial perfusion of an enzymatic, nonionic detergent, ionic detergent, and acid solution with hypotonic and hypertonic rinses was used to systematically remove cellular content. The resultant cardiac ECM retained collagen, elastin, and glycosaminoglycans, and mechanical integrity. Cardiac ECM supported the formation of organized chicken cardiomyocyte sarcomere structure in vitro. The intact decellularized porcine heart provides a tissue engineering template that may be beneficial for future preclinical studies and eventual clinical applications.
authors.affiliation: Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: John M
authors.initials: JM
authors.lastname: Wainwright
authors.forename: Caitlin A
authors.initials: CA
authors.lastname: Czajka
authors.forename: Urvi B
authors.initials: UB
authors.lastname: Patel
authors.forename: Donald O
authors.initials: DO
authors.lastname: Freytes
authors.forename: Kimimasa
authors.initials: K
authors.lastname: Tobita
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Gilbert
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Badylak
mesh_terms: D000818:Animals; D005109:Extracellular Matrix / Q000502:physiology*; D006321:Heart / Q000502:physiology*; D007150:Immunohistochemistry; D008855:Microscopy, Electron, Scanning; D013552:Swine
references.citation: Burns. 2004 Aug;30(5):431-7
references.pmid: 15225907
references.citation: J Bone Joint Surg Am. 2006 Dec;88(12):2665-72
references.pmid: 17142417
references.citation: Cell Motil Cytoskeleton. 1992;21(2):87-100
references.pmid: 1373110
references.citation: Cell Transplant. 2006;15 Suppl 1:S29-40
references.pmid: 16826793
references.citation: J Theor Biol. 1982 Nov 7;99(1):31-68
references.pmid: 6892044
references.citation: J Orthop Res. 2009 Jul;27(7):937-42
references.pmid: 19105224
references.citation: Biomaterials. 2004 May;25(12):2353-61
references.pmid: 14741600
references.citation: Ann Thorac Surg. 2006 Dec;82(6):2050-8
references.pmid: 17126109
references.citation: Tissue Eng. 2007 Mar;13(3):551-66
references.pmid: 17518602
references.citation: J Thorac Cardiovasc Surg. 2007 Apr;133(4):979-85
references.pmid: 17382638
references.citation: Am J Sports Med. 2001 Mar-Apr;29(2):175-84
references.pmid: 11292042
references.citation: J Biomed Mater Res A. 2007 Apr;81(1):250-6
references.pmid: 17269134
references.citation: J Surg Res. 2005 Sep;128(1):87-97
references.pmid: 15922361
references.citation: In Vitro Cell Dev Biol Anim. 2008 Mar-Apr;44(3-4):45-50
references.pmid: 18297366
references.citation: Tissue Eng. 2004 Sep-Oct;10(9-10):1346-58
references.pmid: 15588395
references.citation: J Thorac Cardiovasc Surg. 2003 Dec;126(6):2003-10
references.pmid: 14688719
references.citation: Burns. 1995 Jun;21(4):243-8
references.pmid: 7662122
references.citation: Nat Med. 2008 Feb;14(2):213-21
references.pmid: 18193059
references.citation: J Biomed Mater Res B Appl Biomater. 2008 Feb;84(2):408-14
references.pmid: 17618508
references.citation: J Surg Res. 2008 Jun 1;147(1):61-7
references.pmid: 17950323
references.citation: J Biomed Mater Res A. 2006 Dec 15;79(4):771-8
references.pmid: 16871513
references.citation: Annu Rev Cell Dev Biol. 2006;22:287-309
references.pmid: 16824016
references.citation: Ann Thorac Surg. 2007 Apr;83(4):1501-7
references.pmid: 17383366
references.citation: Arch Intern Med. 2007 May 28;167(10):989-97
references.pmid: 17533201
references.citation: Tissue Eng. 2007 Sep;13(9):2301-10
references.pmid: 17561801
references.citation: Nature. 2008 Feb 21;451(7181):937-42
references.pmid: 18288183
references.citation: J Surg Res. 2009 Mar;152(1):135-9
references.pmid: 18619621
references.citation: Biomaterials. 2006 Jul;27(19):3675-83
references.pmid: 16519932
references.citation: Lancet. 2008 Dec 13;372(9655):2023-30
references.pmid: 19022496
references.citation: Eur J Cardiothorac Surg. 2005 Apr;27(4):566-71
references.pmid: 15784352
references.citation: Tissue Eng Part C Methods. 2009 Jun;15(2):191-200
references.pmid: 19196128
references.citation: J Bone Joint Surg Am. 2007 Mar;89(3):621-30
references.pmid: 17332112
references.citation: Am J Physiol Heart Circ Physiol. 2006 Oct;291(4):H1829-37
references.pmid: 16617136
references.citation: Ann Thorac Surg. 2008 Sep;86(3):967-74; discussion 967-74
references.pmid: 18721593
references.citation: Acta Biomater. 2009 Jan;5(1):1-13
references.pmid: 18938117
references.citation: Hernia. 2009 Apr;13(2):189-99
references.pmid: 19198755
references.citation: J Orthop Res. 2006 Jun;24(6):1299-309
references.pmid: 16649228
references.citation: Biomaterials. 2009 Oct;30(29):5409-16
references.pmid: 19608268