Fabrication of a cell-adhesive protein imprinting surface with an artificial cell membrane structure for cell capturing.
We proposed a new molecular imprinting procedure based on molecular integration for the purpose of cell capture. We selected the cell-adhesive protein fibronectin (FN) as the imprinting protein for preparing templates and evaluated selective cell adhesion on the FN imprinting substrate. Silica beads with a diameter of 15 microm were used as the stamp matrix and FN molecules were adsorbed as a monolayer. The FN recognition sites were constructed by integrating a surfactant as the ligand and immobilizing it with new biocompatible photoreactive phospholipid polymer composed of 2-methacryloyloxyethyl phosphorylcholine (MPC) units. As control substrates, imprinting procedures were carried out using albumin (BSA imprinting substrate) and without imprinting protein (non-imprinting substrate). The binding of FN from the cell culture medium with the fetal calf serum was achieved on the FN imprinting substrate, and induced the cell adhesion. On the other hand, on the non-imprinted and BSA imprinting substrates, the FN scarcely bound from the cell culture medium, and subsequent cell adhesion could not be observed on the substrate. These results indicate that the FN binding sites were well constructed by arranging the ligand surfactant to a suitable position and immobilized by the photoreactive MPC polymer. The MPC polymer prevented the nonspecific adsorption of proteins from the cell culture medium. We concluded that this procedure is convenient and can be potentially used for the preparation of surfaces for cell engineering devices.
Fukazawa
Kyoko
K
Department of Materials Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Ishihara
Kazuhiko
K
D000818:Animals; D001665:Binding Sites; D015374:Biosensing Techniques; D002417:Cattle; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000737:chemistry*; D002460:Cell Line; D005353:Fibronectins / Q000737:chemistry; D066298:In Vitro Techniques; D008567:Membranes, Artificial; D008689:Methacrylates / Q000737:chemistry; D051379:Mice; D008855:Microscopy, Electron, Scanning; D054802:Molecular Imprinting / Q000379:methods*; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012822:Silicon Dioxide; D013499:Surface Properties
D015815:Cell Adhesion Molecules; D005353:Fibronectins; D008567:Membranes, Artificial; D008689:Methacrylates; D011108:Polymers; D010767:Phosphorylcholine; D012710:Serum Albumin, Bovine; C070638:2-methacryloyloxyethyl phosphorylcholine; D012822:Silicon Dioxide