title: Immobilization of cell-adhesive peptides to temperature-responsive surfaces facilitates both serum-free cell adhesion and noninvasive cell harvest.
abstract: We have developed temperature-responsive cell culture surfaces to harvest intact cell sheets for tissue-engineering applications. Both cost and safety issues (e.g., prions, bovine spongiform encephalopathy) are compelling reasons to avoid use of animal-derived materials, including serum, in such culture. In the present study, synthetic cell-adhesive peptides are immobilized onto temperature-responsive polymer-grafted surfaces, and cell adhesion and detachment under serum-free conditions were examined. The temperature-responsive polymer poly(N-isopropylacrylamide) (PI-PAAm) was functionalized by copolymerization with a reactive comonomer having both a carboxyl group and an isopropylacrylamide group. These copolymers were covalently grafted onto tissue culture-grade polystyrene dishes. Synthetic cell-adhesive peptides were then immobilized onto these surfaces via carboxyl groups. Bovine aortic endothelial cells both adhered and spread on these surfaces even under serum-free conditions at 37 degrees C, similar to those in 10% serum-supplemented culture. Spread cells promptly detached from the surfaces on lowering culture temperatures below the lower critical solution temperature of the polymer, 32 degrees C. These surfaces would be useful for serumfree culture for tissue-engineering applications.
authors.affiliation: Department of Applied Chemistry, Waseda University, Tokyo, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Ebara
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamato
authors.forename: Takao
authors.initials: T
authors.lastname: Aoyagi
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kikuchi
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Sakai
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
mesh_terms: D000327:Adsorption; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D018914:Cells, Immobilized / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D016895:Culture Media, Serum-Free / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D008422:Materials Testing; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology*; D013499:Surface Properties; D013552:Swine; D013696:Temperature; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D015815:Cell Adhesion Molecules; D020099:Coated Materials, Biocompatible; D016895:Culture Media, Serum-Free; D009842:Oligopeptides; C046617:arginyl-glycyl-aspartyl-serine