title: Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture.
abstract: We report engineering of half-centimeter-sized bone constructs created in vitro using human adipose-derived stem cells (hASCs), decellularized bone scaffolds, and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4 mm diameter x 4 mm thick), providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400 microm/s), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium-beta-glycerophosphate, and ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, and bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture, and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs.
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, New York, USA.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Fröhlich
authors.forename: Warren L
authors.initials: WL
authors.lastname: Grayson
authors.forename: Darja
authors.initials: D
authors.lastname: Marolt
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Gimble
authors.forename: Nevenka
authors.initials: N
authors.lastname: Kregar-Velikonja
authors.forename: Gordana
authors.initials: G
authors.lastname: Vunjak-Novakovic
mesh_terms: D019149:Bioreactors; D018786:Bone Substitutes; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D006801:Humans; D008297:Male; D010012:Osteogenesis; D010477:Perfusion; D050151:Subcutaneous Fat / Q000166:cytology* / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D018786:Bone Substitutes
references.citation: J Bone Miner Res. 2006 Oct;21(10):1608-17
references.pmid: 16995816
references.citation: J Biomed Mater Res A. 2003 Oct 1;67(1):87-95
references.pmid: 14517865
references.citation: Ann Biomed Eng. 2004 Jan;32(1):112-22
references.pmid: 14964727
references.citation: Biochem Biophys Res Commun. 2008 Jun 6;370(3):456-60
references.pmid: 18395007
references.citation: Tissue Eng. 2004 Mar-Apr;10(3-4):381-91
references.pmid: 15165455
references.citation: Calcif Tissue Int. 1998 Jan;62(1):74-82
references.pmid: 9405737
references.citation: J Biosci Bioeng. 2008 Jun;105(6):586-94
references.pmid: 18640597
references.citation: J Cell Biochem. 2003 May 15;89(2):364-72
references.pmid: 12704799
references.citation: J Cell Biochem. 1997 Dec 1;67(3):386-96
references.pmid: 9361193
references.citation: Stem Cells Dev. 2005 Feb;14(1):92-102
references.pmid: 15725748
references.citation: J Biol Chem. 1989 Oct 5;264(28):16957-62
references.pmid: 2777815
references.citation: J Biomed Mater Res. 2002 Oct;62(1):136-48
references.pmid: 12124795
references.citation: Macromol Biosci. 2004 Aug 9;4(8):743-65
references.pmid: 15468269
references.citation: Biomaterials. 2005 Jun;26(16):3173-85
references.pmid: 15603812
references.citation: J Biomed Mater Res B Appl Biomater. 2006 Jan;76(1):230-9
references.pmid: 16047328
references.citation: Biomaterials. 2000 Dec;21(23):2347-59
references.pmid: 11055282
references.citation: J Biomed Mater Res A. 2006 Jul;78(1):183-93
references.pmid: 16628549
references.citation: Osteoarthritis Cartilage. 2005 Oct;13(10):845-53
references.pmid: 16129630
references.citation: Stem Cells. 2007 Jul;25(7):1610-7
references.pmid: 17395773
references.citation: Tissue Eng. 2001 Apr;7(2):211-28
references.pmid: 11304456
references.citation: Ann Plast Surg. 2006 May;56(5):543-8
references.pmid: 16641633
references.citation: Biomaterials. 2006 Dec;27(36):6138-49
references.pmid: 16895736
references.citation: Plast Reconstr Surg. 2006 Apr 15;117(5):1462-70
references.pmid: 16641714
references.citation: Proc Natl Acad Sci U S A. 2003 Dec 9;100(25):14683-8
references.pmid: 14657343
references.citation: J Biomed Mater Res A. 2008 Jan;84(1):191-7
references.pmid: 17607760
references.citation: J Cell Biochem. 1997 Feb;64(2):278-94
references.pmid: 9027588
references.citation: Stem Cells. 2006 Feb;24(2):376-85
references.pmid: 16322640
references.citation: Cells Tissues Organs. 2008;187(2):89-102
references.pmid: 17938566
references.citation: Biomaterials. 2008 Dec;29(36):4792-9
references.pmid: 18823660
references.citation: Tissue Eng. 2004 Mar-Apr;10(3-4):371-80
references.pmid: 15165454
references.citation: Stem Cells. 2006 May;24(5):1246-53
references.pmid: 16410391
references.citation: Bone. 2005 Nov;37(5):688-98
references.pmid: 16140599
references.citation: J Oral Maxillofac Surg. 2004 May;62(5):601-6
references.pmid: 15122567
references.citation: J Cell Mol Med. 2005 Jan-Mar;9(1):72-84
references.pmid: 15784166
references.citation: Biomaterials. 1998 Aug;19(15):1405-12
references.pmid: 9758040
references.citation: Tissue Eng. 2001 Dec;7(6):729-41
references.pmid: 11749730
references.citation: Tissue Eng. 2005 Nov-Dec;11(11-12):1780-8
references.pmid: 16411823
references.citation: Stem Cells. 2006 Oct;24(10):2262-9
references.pmid: 16794268
references.citation: Iowa Orthop J. 1995;15:118-40
references.pmid: 7634023
references.citation: J Bone Joint Surg Am. 1997 Nov;79(11):1699-709
references.pmid: 9384430
references.citation: Ann Plast Surg. 2003 Jun;50(6):610-7
references.pmid: 12783012
references.citation: Ann Plast Surg. 2006 Jul;57(1):89-93
references.pmid: 16799316
references.citation: Tissue Eng. 2007 Dec;13(12):2981-93
references.pmid: 17916040
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12600-5
references.pmid: 12242339
references.citation: Calcif Tissue Int. 2000 Oct;67(4):314-20
references.pmid: 11000346
references.citation: Am J Physiol Cell Physiol. 2006 Apr;290(4):C1139-46
references.pmid: 16291817
references.citation: J Biomed Mater Res A. 2005 Mar 1;72(3):326-34
references.pmid: 15657936
references.citation: Stem Cells. 2007 Jul;25(7):1823-9
references.pmid: 17446558
references.citation: Nat Biotechnol. 2004 May;22(5):560-7
references.pmid: 15077117
references.citation: J Periodontol. 2000 Aug;71(8):1258-69
references.pmid: 10972641
references.citation: Injury. 2005 Nov;36 Suppl 3:S20-7
references.pmid: 16188545
references.citation: Biomaterials. 2005 Mar;26(9):971-7
references.pmid: 15369685
references.citation: Tissue Eng. 2006 Oct;12(10):2975-83
references.pmid: 17518665
references.citation: Tissue Eng Part A. 2008 Nov;14(11):1809-20
references.pmid: 18620487
references.citation: Biomater Artif Cells Artif Organs. 1990;18(3):383-401
references.pmid: 2383666