Recapitulating cell-cell adhesion using N-cadherin biologically tethered to substrates

Johana C M Vega L., Min Kyung Lee, Jae Hyun Jeong, Cartney E. Smith, Kwan Young Lee, Hee Jung Chung, Deborah E. Leckband, Hyunjoon Kong

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)2172-2179
Number of pages8
JournalBiomacromolecules
Volume15
Issue number6
DOIs
StatePublished - Jun 9 2014

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Cell adhesion
Cadherins
Substrates
Adhesion
Gels
Antibodies
Immunoglobulin Fc Fragments
Glass
Tissue regeneration
Methacrylates
Polyacrylates
Hydrogels
Polyethylene glycols
Lysine
Esters
Polymers
Bone
Proteins
Molecules

ASJC Scopus subject areas

  • Bioengineering
  • Biomaterials
  • Polymers and Plastics
  • Materials Chemistry

Cite this

Recapitulating cell-cell adhesion using N-cadherin biologically tethered to substrates. / 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.

In: Biomacromolecules, Vol. 15, No. 6, 09.06.2014, p. 2172-2179.

Research output: Contribution to journalArticle

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. / Recapitulating cell-cell adhesion using N-cadherin biologically tethered to substrates. In: Biomacromolecules. 2014 ; Vol. 15, No. 6. pp. 2172-2179.
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