title: An adaptable hydrogel array format for 3-dimensional cell culture and analysis.
abstract: Hydrogels have been commonly used as model systems for 3-dimensional (3-D) cell biology, as they have material properties that resemble natural extracellular matrices (ECMs), and their cell-interactive properties can be readily adapted in order to address a particular hypothesis. Natural and synthetic hydrogels have been used to gain fundamental insights into virtually all aspects of cell behavior, including cell adhesion, migration, and differentiated function. However, cell responses to complex 3-D environments are difficult to adequately explore due to the large number of variables that must be controlled simultaneously. Here we describe an adaptable, automated approach for 3-D cell culture within hydrogel arrays. Our initial results demonstrate that the hydrogel network chemistry (both natural and synthetic), cell type, cell density, cell adhesion ligand density, and degradability within each array spot can be systematically varied to screen for environments that promote cell viability in a 3-D context. In a test-bed application we then demonstrate that a hydrogel array format can be used to identify environments that promote viability of HL-1 cardiomyocytes, a cell line that has not been cultured previously in 3-D hydrogel matrices. Results demonstrate that the fibronectin-derived cell adhesion ligand RGDSP improves HL-1 viability in a dose-dependent manner, and that the effect of RGDSP is particularly pronounced in degrading hydrogel arrays. Importantly, in the presence of 70mum RGDSP, HL-1 cardiomyocyte viability does not decrease even after 7 days of culture in PEG hydrogels. Taken together, our results indicate that the adaptable, array-based format developed in this study may be useful as an enhanced throughput platform for 3-D culture of a variety of cell types.
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53706, United States.
authors.forename: Leenaporn
authors.initials: L
authors.lastname: Jongpaiboonkit
authors.forename: William J
authors.initials: WJ
authors.lastname: King
authors.forename: Gary E
authors.initials: GE
authors.lastname: Lyons
authors.forename: Amy L
authors.initials: AL
authors.lastname: Paguirigan
authors.forename: Jay W
authors.initials: JW
authors.lastname: Warrick
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
authors.forename: William L
authors.initials: WL
authors.lastname: Murphy
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002448:Cell Adhesion; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002460:Cell Line; D002478:Cells, Cultured; D024042:Collagen Type I / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology; D041681:NIH 3T3 Cells; D011092:Polyethylene Glycols / Q000737:chemistry
chemical_list: D001672:Biocompatible Materials; D024042:Collagen Type I; D003470:Culture Media; D020100:Hydrogels; D011092:Polyethylene Glycols