title: Culture of HepG2 liver cells on three dimensional polystyrene scaffolds enhances cell structure and function during toxicological challenge.
abstract: Cultured cells are dramatically affected by the micro-environment in which they are grown. In this study, we have investigated whether HepG2 liver cells grown in three dimensional (3-D) cultures cope more effectively with the known cytotoxic agent, methotrexate, than their counterparts grown on traditional two dimensional (2-D) flat plastic surfaces. To enable 3-D growth of HepG2 cells in vitro, we cultured cells on 3-D porous polystyrene scaffolds previously developed in our laboratories. HepG2 cells grown in 3-D displayed excellent morphological characteristics and formed numerous bile canaliculi that were seldom seen in cultures grown on 2-D surfaces. The function of liver cells grown on 3-D supports was significantly enhanced compared to activity of cells grown on 2-D standard plasticware. Unlike their 2-D counterparts, 3-D cultures were less susceptible to lower concentrations of methotrexate. Cells grown in 3-D maintained their structural integrity, possessed greater viability, were less susceptible to cell death at higher levels of the cytotoxin compared to 2-D cultures, and appeared to respond to the drug in a manner more comparable to its known activity in vivo. Our results suggest that hepatotoxicity testing using 3-D cultures might be more likely to reflect true physiological responses to cytotoxic compounds than existing models that rely on 2-D culture systems. This technology has potential applications for toxicity testing and drug screening.
authors: School of Biological and Biomedical Science, Durham University, South Road, Durham DH1 3LE, UK.
authors: Maria
authors: M
authors: Bokhari
authors: Ross J
authors: RJ
authors: Carnachan
authors: Neil R
authors: NR
authors: Cameron
authors: Stefan A
authors: SA
authors: Przyborski
mesh_terms: D002448:Cell Adhesion; D002450:Cell Communication; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D021621:Imaging, Three-Dimensional; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D008727:Methotrexate / Q000633:toxicity; D008854:Microscopy, Electron; D011137:Polystyrenes; D018675:Toxicity Tests / Q000379:methods
chemical_list: D011137:Polystyrenes; D008727:Methotrexate
references: Cell Tissue Res. 1991 Feb;263(2):227-35
references: 1672508
references: Mol Biol Cell. 1993 Jan;4(1):59-69
references: 8443410
references: J Gastroenterol Hepatol. 1998 Sep;13 Suppl:S55-61
references: 9792035
references: J Cell Sci. 2003 Jun 15;116(Pt 12):2377-88
references: 12766184
references: Biochem Biophys Res Commun. 2007 Mar 23;354(4):1095-100
references: 17276400
references: Soft Matter. 2006 Jun 20;2(7):608-616
references: 32680240
references: Exp Cell Res. 2002 Mar 10;274(1):56-67
references: 11855857
references: J Cell Biol. 1997 Apr 7;137(1):231-45
references: 9105051
references: Toxicol In Vitro. 2000 Oct;14(5):475-85
references: 10963964
references: Exp Cell Res. 1990 Feb;186(2):227-35
references: 2298241
references: J Biotechnol. 2006 Apr 10;122(3):372-81
references: 16446003
references: Drug Discov Today. 2002 Jun 1;7(11):612-20
references: 12047872
references: Anal Chem. 2006 Jul 1;78(13):4291-8
references: 16808435
references: Proc Biol Sci. 1997 May 22;264(1382):657-61
references: 9178537
references: Biomed Res. 2006 Apr;27(2):75-80
references: 16707846
references: In Vitro Cell Dev Biol. 1989 Mar;25(3 Pt 1):267-75
references: 2466823
references: Org Lett. 2002 Jul 25;4(15):2497-500
references: 12123360
references: Science. 2001 Nov 23;294(5547):1708-12
references: 11721053
references: J Biochem Biophys Methods. 2005 Mar 31;62(3):231-40
references: 15733583
references: In Vitro Cell Dev Biol Anim. 1996 Apr;32(4):197-203
references: 8727043
references: Tissue Eng. 1999 Aug;5(4):291-304
references: 10477852
references: Inflamm Res. 2005 Jul;54(7):273-80
references: 16134056
references: Methods Enzymol. 2000;322:433-72
references: 10914039
references: Proc Natl Acad Sci U S A. 1983 May;80(10):3078-80
references: 6190170
references: Biomaterials. 2000 Mar;21(5):431-40
references: 10674807
references: J Cell Biol. 1985 Sep;101(3):914-23
references: 2411740
references: J Immunol Methods. 1983 Dec 16;65(1-2):55-63
references: 6606682
references: Biomaterials. 2004 Aug;25(18):3991-4000
references: 15046889
references: Soft Matter. 2005 Jun 27;1(2):107-113
references: 32646082
references: J Biol Chem. 1971 Jun 10;246(11):3500-7
references: 5578905
references: FASEB J. 1999 Dec;13(15):2214-24
references: 10593869
references: J Biomater Sci Polym Ed. 1998;9(10):1049-69
references: 9806445
references: Exp Cell Res. 1992 Oct;202(2):423-31
references: 1327854
references: Biomaterials. 2000 Apr;21(8):783-93
references: 10721747
references: Toxicology. 2004 May 20;198(1-3):329-40
references: 15138059
references: Proc Natl Acad Sci U S A. 2003 Oct 28;100(22):12741-6
references: 14561891
references: Biochim Biophys Acta. 1966 Aug 10;122(2):244-64
references: 5969301
references: Microsc Res Tech. 1992 Jun 1;21(4):271-82
references: 1379091
references: Osteoarthritis Cartilage. 2005 Jun;13(6):527-36
references: 15922187
references: Biomaterials. 2005 Jun;26(17):3673-82
references: 15621258
references: J Am Coll Nutr. 1984;3(1):93-6
references: 6201520
references: Proc Natl Acad Sci U S A. 2004 Dec 28;101(52):18024-9
references: 15601776
references: J Cell Sci. 1988 May;90 ( Pt 1):79-92
references: 3058731
references: Ann Pharmacother. 2002 Sep;36(9):1344-50
references: 12196049
references: Biochem Biophys Res Commun. 2004 Feb 6;314(2):483-8
references: 14733931
references: J Biomed Mater Res A. 2004 Nov 1;71(2):268-74
references: 15368220
references: FASEB J. 1996 Nov;10(13):1471-84
references: 8940293
references: Biomaterials. 1993 Apr;14(5):323-30
references: 8507774
references: Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14821-6
references: 9843973