Culture of human vascular endothelial cells on an RGD-containing synthetic peptide attached to a starch-coated polystyrene surface: comparison with fibronectin-coated tissue grade polystyrene.
A synthetic peptide, Gly-Arg-Gly-Asp-Ser-Pro-Lys (GRGDSPK), which includes the cell-adhesive region of fibronectin, Arg-Gly-Asp (RGD), was covalently bound to a dialdehyde starch (DAS) coating on a polymer surface by reductive amination. The GRGDSPK/DAS-coated surface was characterized by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). AFM and SEM revealed a uniform, roughened, textured surface, much more so than standard polymer or adhesive protein-coated polymer surfaces. XPS showed that GRGDSPK binding to DAS occurred in dose-dependent fashion in the 0-200 micrograms ml-1 GRGDSPK concentration range, with a plateau happening in the 200-400 micrograms ml-1 range. AFM revealed a uniform peptide layer on the DAS surface with a maximum separation distance of 50 nm between peptides. Angle-dependent XPS showed that the peptide is present in nearly constant amounts to at least 10 nm depth of the DAS coating. The attachment, spreading and growth properties of anchorage-dependent human umbilical vein endothelial cells (EC) on the GRGDSPK/DAS-coated polystyrene surface were compared with a standard fibronectin-coated polystyrene surface. EC adhesion, spreading and growth properties were similar for cells plated on polystyrene surfaces coated with fibronectin (5 micrograms cm-2) and GRGDSPK (25-50 micrograms ml-1)/DAS. In contrast, EC adhesion, spreading and growth performance significantly increased for cells plated on GRGDSPK (100-200 micrograms ml-1)/DAS compared with the fibronectin-coated surface. These findings support the conclusion that the GRGDSPK/DAS-coated surface can be substituted for an adhesive protein-coated surface in the culture of anchorage-dependent cells.
Holland
J
J
Department of Medicine, State University of New York Health Science Center, Syracuse 13210, USA.
Hersh
L
L
Bryhan
M
M
Onyiriuka
E
E
Ziegler
L
L
D000595:Amino Acid Sequence; D002448:Cell Adhesion; D002455:Cell Division; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D005353:Fibronectins; D006801:Humans; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D009842:Oligopeptides; D011137:Polystyrenes; D013052:Spectrometry, X-Ray Emission
D005353:Fibronectins; D009842:Oligopeptides; D011137:Polystyrenes; C086456:glycyl- arginyl-glycyl-aspartyl-seryl-prolyl-lysine; C047981:arginyl-glycyl-aspartic acid