title: Fabrication and selective functionalization of amine-reactive polymer multilayers on topographically patterned microwell cell culture arrays.
abstract: We report an approach to the fabrication and selective functionalization of amine-reactive polymer multilayers on the surfaces of 3-D polyurethane-based microwell cell culture arrays. "Reactive" layer-by-layer assembly of multilayers using branched polyethyleneimine (BPEI) and the azlactone-functionalized polymer poly(2-vinyl-4,4'-dimethylazlactone) (PVDMA) yielded film-coated microwell arrays that could be chemically functionalized postfabrication by treatment with different amine-functionalized macromolecules or small molecule primary amines. Treatment of film-coated arrays with the small molecule amine d-glucamine resulted in microwell surfaces that resisted the adhesion and proliferation of mammalian fibroblast cells in vitro. These and other experiments demonstrated that it was possible to functionalize different structural features of these arrays in a spatially resolved manner to create dual-functionalized substrates (e.g., to create arrays having either (i) azlactone-functionalized wells, with regions between the wells functionalized with glucamine or (ii) substrates with spatially resolved regions of two different cationic polymers). In particular, spatial control over glucamine functionalization yielded 3-D substrates that could be used to confine cell attachment and growth to microwells for periods of up to 28 days and support the 3-D culture of arrays of cuboidal cell clusters. These approaches to dual functionalization could prove useful for the long-term culture and maintenance of cell types for which the presentation of specific and chemically well-defined 3-D culture environments is required for control over cell growth, differentiation, and other important behaviors. More generally, our approach provides methods for the straightforward chemical functionalization of otherwise unreactive topographically patterned substrates that could prove to be useful in a range of other fundamental and applied contexts.
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin - Madison, Madison, WI 53706, USA.
authors.forename: Adam H
authors.initials: AH
authors.lastname: Broderick
authors.forename: Samira M
authors.initials: SM
authors.lastname: Azarin
authors.forename: Maren E
authors.initials: ME
authors.lastname: Buck
authors.forename: Sean P
authors.initials: SP
authors.lastname: Palecek
authors.forename: David M
authors.initials: DM
authors.lastname: Lynn
mesh_terms: D000588:Amines / Q000737:chemistry* / Q000378:metabolism; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D020099:Coated Materials, Biocompatible / Q000737:chemistry* / Q000378:metabolism; D005453:Fluorescence; D057809:HEK293 Cells; D006801:Humans; D007783:Lactones / Q000737:chemistry* / Q000378:metabolism; D011094:Polyethyleneimine / Q000737:chemistry* / Q000378:metabolism; D011140:Polyurethanes / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D000588:Amines; D020099:Coated Materials, Biocompatible; D007783:Lactones; D011140:Polyurethanes; D011094:Polyethyleneimine