title: Microfabricated polymeric vessel mimetics for 3-D cancer cell culture.
abstract: Modeling tumor growth in vitro is essential for cost-effective testing of hypotheses in preclinical cancer research. 3-D cell culture offers an improvement over monolayer culture for studying cellular processes in cancer biology because of the preservation of cell-cell and cell-ECM interactions. Oxygen transport poses a major barrier to mimicking in vivo environments and is not replicated in conventional cell culture systems. We hypothesized that we can better mimic the tumor microenvironment using a bioreactor system for controlling gas exchange in cancer cell cultures with silicone hydrogel synthetic vessels. Soft-lithography techniques were used to fabricate oxygen-permeable silicone hydrogel membranes containing arrays of micropillars. These membranes were inserted into a bioreactor and surrounded by basement membrane extract (BME) within which fluorescent ovarian cancer (OVCAR8) cells were cultured. Cell clusters oxygenated by synthetic vessels showed a ∼100μm drop-off to anoxia, consistent with in vivo studies of tumor nodules fed by the microvasculature. Oxygen transport in the bioreactor system was characterized by experimental testing with a dissolved oxygen probe and finite element modeling of convective flow. Our study demonstrates differing growth patterns associated with controlling gas distributions to better mimic in vivo conditions.
authors.affiliation: Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Ashley A
authors.initials: AA
authors.lastname: Jaeger
authors.forename: Chandan K
authors.initials: CK
authors.lastname: Das
authors.forename: Nicole Y
authors.initials: NY
authors.lastname: Morgan
authors.forename: Randall H
authors.initials: RH
authors.lastname: Pursley
authors.forename: Philip G
authors.initials: PG
authors.lastname: McQueen
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Hall
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Pohida
authors.forename: Michael M
authors.initials: MM
authors.lastname: Gottesman
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D055616:Microtechnology; D011108:Polymers / Q000737:chemistry*
chemical_list: D011108:Polymers
keywords: Bioreactor; Carcinogenesis; Hydrogel; Microstructure; Oxygenation; Silicone
references.citation: Mol Oncol. 2007 Jun;1(1):84-96
references.pmid: 18516279
references.citation: Artif Organs. 2000 Apr;24(4):278-88
references.pmid: 10816201
references.citation: Cancer Cell. 2012 Mar 20;21(3):309-22
references.pmid: 22439926
references.citation: Semin Cancer Biol. 2005 Oct;15(5):378-86
references.pmid: 15975825
references.citation: Stem Cells. 2010 Jun;28(6):1019-29
references.pmid: 20506127
references.citation: J Natl Cancer Inst. 2007 Oct 3;99(19):1441-54
references.pmid: 17895480
references.citation: FASEB J. 2006 Dec;20(14):2531-3
references.pmid: 17077286
references.citation: Assay Drug Dev Technol. 2009 Jun;7(3):233-49
references.pmid: 19548831
references.citation: Nat Rev Cancer. 2005 Sep;5(9):675-88
references.pmid: 16148884
references.citation: Biomed Microdevices. 2007 Apr;9(2):123-34
references.pmid: 17160707
references.citation: Nat Rev Cancer. 2006 Aug;6(8):583-92
references.pmid: 16862189
references.citation: Adv Biochem Eng Biotechnol. 2009;112:1-27
references.pmid: 19290495
references.citation: Nat Rev Mol Cell Biol. 2006 Mar;7(3):211-24
references.pmid: 16496023
references.citation: Lab Chip. 2008 Jul;8(7):1042-7
references.pmid: 18584077
references.citation: Integr Biol (Camb). 2013 Mar;5(3):597-605
references.pmid: 23388834
references.citation: Biotechnol J. 2008 Oct;3(9-10):1172-84
references.pmid: 18566957
references.citation: Curr Opin Biotechnol. 2003 Oct;14(5):526-32
references.pmid: 14580584
references.citation: Biomed Microdevices. 2010 Jun;12(3):465-75
references.pmid: 20174871
references.citation: Nanomedicine (Lond). 2010 Apr;5(3):469-84
references.pmid: 20394538
references.citation: Anticancer Res. 2007 May-Jun;27(3A):1295-300
references.pmid: 17593622
references.citation: J Theor Biol. 1982 Nov 7;99(1):31-68
references.pmid: 6892044
references.citation: Cell Biol Int. 1999;23(3):157-61
references.pmid: 10562436
references.citation: Proc Natl Acad Sci U S A. 2011 Nov 15;108(46):18708-13
references.pmid: 22068913
references.citation: Cancer Res. 1985 Sep;45(9):4200-5
references.pmid: 4028010
references.citation: Matrix Biol. 2008 Jul;27(6):573-585
references.pmid: 18411046
references.citation: Theor Biol Med Model. 2009 Apr 16;6:5
references.pmid: 19371422
references.citation: Biomaterials. 2011 Nov;32(31):7905-12
references.pmid: 21782234
references.citation: Biomaterials. 2012 Nov;33(33):8430-41
references.pmid: 22940219
references.citation: Biorheology. 2009;46(5):417-37
references.pmid: 19940357
references.citation: J Biomater Sci Polym Ed. 2011;22(11):1509-22
references.pmid: 20626957
references.citation: J Cell Biol. 1972 Sep;54(3):626-37
references.pmid: 4339818
references.citation: Biomaterials. 2007 Dec;28(36):5487-97
references.pmid: 17881050
references.citation: Cell. 2007 Aug 24;130(4):601-10
references.pmid: 17719539
references.citation: Annu Rev Cell Dev Biol. 2006;22:287-309
references.pmid: 16824016
references.citation: Semin Cancer Biol. 2008 Oct;18(5):311-21
references.pmid: 18455428
references.citation: Biochemistry. 1982 Nov 23;21(24):6188-93
references.pmid: 6217835
references.citation: J Neurosci Methods. 1991 Apr;37(2):173-82
references.pmid: 1715499
references.citation: Br J Cancer. 1968 Jun;22(2):258-73
references.pmid: 5660132
references.citation: Nat Biotechnol. 2004 Feb;22(2):151-2
references.pmid: 14755282
references.citation: Nat Methods. 2007 Oct;4(10):855-60
references.pmid: 17767164
references.citation: J Natl Cancer Inst. 1994 Dec 21;86(24):1846-52
references.pmid: 7990159
references.citation: Biotechnol Bioeng. 2007 Jan 1;96(1):156-66
references.pmid: 16917927
references.citation: Biomaterials. 2001 Dec;22(24):3273-83
references.pmid: 11700799
references.citation: Life Sci. 1995;57(2):131-41
references.pmid: 7603295
references.citation: Acta Biomater. 2011 Sep;7(9):3312-24
references.pmid: 21704736
references.citation: J Cell Biol. 1997 Apr 7;137(1):231-45
references.pmid: 9105051
references.citation: Int J Radiat Biol. 2007 Nov-Dec;83(11-12):849-71
references.pmid: 18058370
references.citation: Tissue Eng Part C Methods. 2012 Jan;18(1):54-61
references.pmid: 21902468
references.citation: J Cell Physiol. 2009 Oct;221(1):18-25
references.pmid: 19492404
references.citation: Science. 1988 Apr 8;240(4849):177-84
references.pmid: 2451290
references.citation: Biotechnol Bioeng. 2008 Apr 15;99(6):1472-81
references.pmid: 17969156
references.citation: Nat Rev Mol Cell Biol. 2007 Oct;8(10):839-45
references.pmid: 17684528
references.citation: Nat Mater. 2010 Feb;9(2):90-3
references.pmid: 20094076
references.citation: Cell Mol Bioeng. 2012 Sep 1;5(3):299-306
references.pmid: 23730348
references.citation: Anal Chem. 2003 Dec 1;75(23):6544-54
references.pmid: 14640726
references.citation: J Natl Cancer Inst. 2013 Apr 3;105(7):452-8
references.pmid: 23434901
references.citation: Trends Biotechnol. 2004 Feb;22(2):80-6
references.pmid: 14757042
references.citation: Br J Cancer. 1955 Dec;9(4):539-49
references.pmid: 13304213
references.citation: Biomaterials. 2008 May;29(14):2259-69
references.pmid: 18289662
references.citation: J Biotechnol. 2010 Jul 1;148(1):3-15
references.pmid: 20097238
references.citation: Biomaterials. 2010 Nov;31(32):8494-506
references.pmid: 20709389
references.citation: J Cell Physiol. 1992 May;151(2):386-94
references.pmid: 1572910
references.citation: J Surg Res. 1992 Dec;53(6):549-57
references.pmid: 1494286
references.citation: Biomaterials. 2011 Sep;32(26):6052-8
references.pmid: 21640378