Recapitulating Cell-Cell Adhesion Using N-Cadherin Biologically Tethered to Substrates
- Authors
- Vega L, Johana C. M.; Lee, Min Kyung; Jeong, Jae Hyun; Smith, Cartney E.; Lee, Kwan Young; Chung, Hee Jung; Leckband, Deborah E.; Kong, Hyunjoon
- Issue Date
- Jun-2014
- Publisher
- AMER CHEMICAL SOC
- Citation
- BIOMACROMOLECULES, v.15, no.6, pp.2172 - 2179
- Journal Title
- BIOMACROMOLECULES
- Volume
- 15
- Number
- 6
- Start Page
- 2172
- End Page
- 2179
- URI
- http://scholarworks.bwise.kr/ssu/handle/2018.sw.ssu/10034
- DOI
- 10.1021/bm500335w
- ISSN
- 1525-7797
- Abstract
- Intercellular adhesion modulated by cadherin molecules plays an important role in diverse cellular functions including tissue morphogenesis, regeneration, and pathogenesis. However, it is a challenging task to decipher the effects of cell cell adhesion in vitro because of difficulty in controlling the extent and numbers of cell cell contacts. In this study, we hypothesize that tethering recombinant extracellular domains of neural cadherin with a C-terminal immunoglobulin Fc domain (N-Cad-Fc) to a substrate with an immobilized anti-Fc antibody (Fc-antibody) and a bifunctional polymer, which is reactive to both protein and substrate, would allow us to recapitulate cell cell adhesion, independent of the number of cells plated on the substrate. To examine this hypothesis, we first immobilized Fc-antibody to a polyacrylamide hydrogel and a methacrylate-substituted glass using poly(amino-2-hydroxyethyl-co-2-methacryloxyethyl aspartamide)-g-poly(ethylene glycol)-N-hydroxysuccinimide ester (PHMAA-g-PEGNHS) and then incubated the gel in medium containing defined concentrations of the recombinant N-Cad-Fc. The resulting N-Cad-conjugated substrate enabled us to modulate adhesion of bone marrow stromal cells to the gel surface by varying the surface density of N-Cad-Fc. In contrast, direct chemical conjugation of N-Cad-Fc to the gel surface did not support cell adhesion. Additionally, the glass substrate biologically tethered with N-Cad-Fc promoted neuronal adhesion significantly more than substrates coated with poly-L-lysine. We suggest that this novel biological tethering method could be broadly applicable for modifying substrates with a variety of classical cadherins to enable the systematic study of the effects of cadherin-modulated cell cell adhesion on cellular activities.
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