Bioconjugafe Chem. 1995, 6, 41 1-41 7
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Photoimmobilization of a Bioactive Laminin Fragment and Pattern-Guided Selective Neuronal Cell Attachment Jean-FranCois C16mence,t J o h n P. Ranieri,$ Patrick Aebischer,$ a n d Hans Sigrist",' Institute of Biochemistry, University of Bern, Freiestrasse 3, CH-3012 Berne, Switzerland, and Division of Surgical Research, Centre Hospitalier Universitaire Vaudois, CH-1011 Lausanne, Switzerland. Received December 29, 1994@
To attain light-dependent functionalization of biocompatible materials, a photolabel-derivatized, bioactive laminin fragment has been synthesized, chemically characterized, and photoimmobilized. Covalent high-resolution patterning of the laminin fragment CDPGYIGSR to hydroxylated fluorinated ethylene propylene (FEP-OH), poly(viny1 alcohol), and glycophase glass has been achieved. The synthetic peptide CDPGYIGSR was thermochemically coupled to either N-[m-[3-(trifluoromethyl)diazirin-3-yllphenyll-4-maleimidobutyramide or 4-maleimidobenzophenone. Photolabel-derivatized peptides were radiolabeled, and 20 and 300 pm-sized patterns were visualized by autoradiography. The biospecific interaction of photoimmobilized laminin fragments with cells was investigated by analyzing the selective attachment of NG 108-15 neuroblastoma x glioma cells which bear CDPGYIGSR-specific cell surface receptors. On photopatterned FEP-OH membranes NG 108-15 cells differentiated in serum-supplemented media within 1 day. Specific attachment to the immobilized oligopeptide CDPGYIGSR was assessed in serum-free media with competitive binding studies, showing an 82% decrease in cell adherence after the cell receptors were blocked with soluble CDPGYIGSR.
al., 1993). Topically selective cell attachment has been reported to occur with physically adsorbed extracellular Tissue-engineered material systems require the momatrix proteins (e.g., laminin, fibronectin) and glycoslecular design of biomaterial5 to elicit a desired, receptoraminoglycans such as keratan sulfate and chondroitin specific cellular response (Aebischer et al., 1992; Peppas sulfate (Snow et al., 1990). Fromherz et al. (1991) have and Langer, 1994; Ratner, 1993; Bellamkonda and Aeaccomplished the same goal by irradiating physically bischer, 1994). Two-dimensional patterned substrates adsorbed laminin through a mask using UV radiation have been developed to control neural cell attachment
