title: Bio-functionalized thermoresponsive interfaces facilitating cell adhesion and proliferation.
abstract: Bio-functionalized thermoresponsive culture interfaces co-immobilized with cell adhesive peptide, RGDS, and cell growth factor, insulin (INS), are investigated to promote initial cell adhesion and cell growth for further cell sheet engineering applications. These bio-functionalized interfaces were prepared by electron beam-induced copolymerization of N-isopropylacrylamide (IPAAm) with its carboxyl-derivatized analog, 2-carboxyisopropylacrylamide (CIPAAm), and grafting onto tissue culture polystyrene dishes, followed by immobilization of RGDS and/or INS to CIPAAm carboxyls. Adhesion and proliferation of bovine carotid artery endothelial cells (ECs) were examined on the RGDS-INS co-immobilized thermoresponsive interfaces. Immobilized RGDS facilitated initial EC adhesion on the surfaces and INS modification was demonstrated to induce EC proliferation, respectively. More pronounced EC growth was indicated by co-immobilization of appropriate amount of RGDS and INS. This may be due to synergistic effect of direct co-stimulation of adhered ECs by surface-immobilized RGDS and INS molecules. ECs grown on the RGDS-INS co-immobilized thermoresponsive interfaces can also be recovered spontaneously as viable tissue monolayers by solely reducing culture temperature. RGDS-INS co-immobilized thermoresponsive interfaces strongly supported initial EC adhesion and growth than unmodified thermoresponsive surfaces even under serum-free culture. Addition of soluble growth factors to serum-free culture medium effectively induced EC proliferation to confluency. Co-immobilization of cell adhesion peptides and growth factors on thermoresponsive surfaces should be effective for rapid preparation of intact cell sheets and their utilization to regenerative medicine.
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Center of Excellence (COE) Program for the 21st Century, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Hatakeyama
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kikuchi
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamato
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
mesh_terms: D000178:Acrylamides; D000818:Animals; D001672:Biocompatible Materials; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology; D006801:Humans; D007328:Insulin; D009842:Oligopeptides; D011137:Polystyrenes
chemical_list: C404843:2-carboxyisopropylacrylamide; D000178:Acrylamides; D001672:Biocompatible Materials; D007328:Insulin; D009842:Oligopeptides; D011137:Polystyrenes; C046617:arginyl-glycyl-aspartyl-serine; C067295:N-isopropylacrylamide