title: Long term perfusion system supporting adipogenesis.
abstract: Adipose tissue engineered models are needed to enhance our understanding of disease mechanisms and for soft tissue regenerative strategies. Perfusion systems generate more physiologically relevant and sustainable adipose tissue models, however adipocytes have unique properties that make culturing them in a perfusion environment challenging. In this paper we describe the methods involved in the development of two perfusion culture systems (2D and 3D) to test their applicability for long term in vitro adipogenic cultures. It was hypothesized that a silk protein biomaterial scaffold would provide a 3D framework, in combination with perfusion flow, to generate a more physiologically relevant sustainable adipose tissue engineered model than 2D cell culture. Consistent with other studies evaluating 2D and 3D culture systems for adipogenesis we found that both systems successfully model adipogenesis, however 3D culture systems were more robust, providing the mechanical structure required to contain the large, fragile adipocytes that were lost in 2D perfused culture systems. 3D perfusion also stimulated greater lipogenesis and lipolysis and resulted in decreased secretion of LDH compared to 2D perfusion. Regardless of culture configuration (2D or 3D) greater glycerol was secreted with the increased nutritional supply provided by perfusion of fresh media. These results are promising for adipose tissue engineering applications including long term cultures for studying disease mechanisms and regenerative approaches, where both acute (days to weeks) and chronic (weeks to months) cultivation are critical for useful insight.
authors.affiliation: Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States.
authors.forename: Rosalyn D
authors.initials: RD
authors.lastname: Abbott
authors.affiliation: Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States; Department of Brain and Cognitive Science, MIT, Cambridge, MA, United States.
authors.forename: Waseem K
authors.initials: WK
authors.lastname: Raja
authors.affiliation: Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States.
authors.forename: Rebecca Y
authors.initials: RY
authors.lastname: Wang
authors.affiliation: Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States.
authors.forename: Jordan A
authors.initials: JA
authors.lastname: Stinson
authors.affiliation: Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States; Biological Engineering, MIT, Cambridge, MA, United States.
authors.forename: Dean L
authors.initials: DL
authors.lastname: Glettig
authors.affiliation: Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States; Department of Chemical and Biomolecular Engineering, University of Connecticut, Storrs, CT, United States.
authors.forename: Kelly A
authors.initials: KA
authors.lastname: Burke
authors.affiliation: Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States. Electronic address: david.kaplan@tufts.edu.
authors.forename: David L
authors.initials: DL
authors.lastname: Kaplan
mesh_terms: D017667:Adipocytes / Q000166:cytology; D050156:Adipogenesis / Q000502:physiology*; D053687:Adult Stem Cells / Q000166:cytology; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D006801:Humans; D008422:Materials Testing; D010477:Perfusion / Q000295:instrumentation; D047011:Silk / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D001672:Biocompatible Materials; D047011:Silk
keywords: Adipose tissue engineering; Long term culture; Perfusion; Silk scaffold; Three dimensional culture
references.citation: Biomaterials. 2002 Oct;23(20):4131-41
references.pmid: 12182315
references.citation: J Extra Corpor Technol. 2003 Sep;35(3):200-2
references.pmid: 14653420
references.citation: Curr Stem Cell Res Ther. 2012 Jul;7(4):287-92
references.pmid: 22563665
references.citation: Cytotechnology. 1993;11(3):189-95
references.pmid: 7764124
references.citation: PLoS One. 2013;8(2):e55696
references.pmid: 23405199
references.citation: Biomatter. 2012 Oct-Dec;2(4):167-75
references.pmid: 23507883
references.citation: Biotechnol Prog. 2004 May-Jun;20(3):905-12
references.pmid: 15176898
references.citation: Lab Chip. 2009 Nov 21;9(22):3185-92
references.pmid: 19865724
references.citation: Trends Biotechnol. 2004 Feb;22(2):80-6
references.pmid: 14757042
references.citation: Biomaterials. 2013 Apr;34(12):2960-8
references.pmid: 23374707
references.citation: Biomaterials. 2005 Jan;26(2):147-55
references.pmid: 15207461
references.citation: Exp Hematol. 2005 Nov;33(11):1402-16
references.pmid: 16263424
references.citation: Differentiation. 2007 Jun;75(5):360-70
references.pmid: 17286602
references.citation: Tissue Eng Part B Rev. 2012 Feb;18(1):62-75
references.pmid: 21902622
references.citation: Anal Bioanal Chem. 2010 Aug;397(7):2939-47
references.pmid: 20549489
references.citation: Tissue Eng Part C Methods. 2012 Jan;18(1):54-61
references.pmid: 21902468
references.citation: Folia Biol (Praha). 2011;57(2):47-56
references.pmid: 21631961
references.citation: Tissue Eng Part A. 2009 Aug;15(8):2227-36
references.pmid: 19207036
references.citation: Anal Chem. 2009 Mar 15;81(6):2350-6
references.pmid: 19231843
references.citation: J Clin Endocrinol Metab. 2008 Nov;93(11 Suppl 1):S57-63
references.pmid: 18987271
references.citation: Cell. 2006 May 5;125(3):577-91
references.pmid: 16678100
references.citation: Biomaterials. 2006 Dec;27(36):6052-63
references.pmid: 16973213
references.citation: Tissue Eng Part C Methods. 2013 Oct;19(10):745-54
references.pmid: 23373822
references.citation: Nat Protoc. 2011 Oct;6(10):1612-31
references.pmid: 21959241
references.citation: Curr Opin Cell Biol. 2002 Oct;14(5):633-9
references.pmid: 12231360
references.citation: Anat Rec. 2001 Aug 1;263(4):361-6
references.pmid: 11500812
references.citation: Tissue Eng Part A. 2011 May;17(9-10):1437-44
references.pmid: 21247356
references.citation: J Diabetes Sci Technol. 2014 Nov;8(6):1227-32
references.pmid: 25160666
references.citation: Methods Mol Biol. 2008;449:69-79
references.pmid: 18370084
references.citation: Biomaterials. 2006 Dec;27(36):6064-82
references.pmid: 16890988
references.citation: Tissue Eng Part A. 2009 Nov;15(11):3389-99
references.pmid: 19402786
references.citation: Open Orthop J. 2011;5 Suppl 2:267-70
references.pmid: 21886691
references.citation: Tissue Eng Part C Methods. 2012 Dec;18(12):958-67
references.pmid: 22651694
references.citation: Tissue Eng Part C Methods. 2010 Oct;16(5):1157-65
references.pmid: 20144013
references.citation: Sci Rep. 2012;2:799
references.pmid: 23150771
references.citation: Biotechnol Prog. 2004 Mar-Apr;20(2):590-7
references.pmid: 15059006