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Bi-ligand surfaces with oriented and patterned protein for real-time tracking of cell migration.


ABSTRACT: A bioactive platform for the quantitative observation of cell migration is presented by (1) presenting migration factors in a well-defined manner on 2-D substrates, and (2) enabling continuous cell tracking. Well-defined substrate presentation is achieved by correctly orienting immobilized proteins (chemokines and cell adhesion molecules), such that the active site is accessible to cell surface receptors. A thiol-terminated self-assembled monolayer on a silica slide was used as a base substrate for subsequent chemistry. The thiol-terminated surface was converted to an immobilized metal ion surface using a maleimido-nitrilotriacetic acid (NTA) cross-linker that bound Histidine-tagged recombinant proteins on the surface with uniform distribution and specific orientation. This platform was used to study the influence of surface-immobilized chemokine SDF-1? and cell adhesion molecule ICAM-1 on murine splenic B lymphocyte migration. While soluble SDF-1? induced trans-migration in a Boyden Chamber type chemotaxis assay, immobilized SDF-1? alone did not elicit significant surface-migration on our test-platform surface. Surface-immobilized cell adhesion protein, ICAM-1, in conjunction with activation enabled migration of this cell type on our surface. Controlled exposure to UV light was used to produce stable linear gradients of His-tagged recombinant SDF-1? co-immobilized with ICAM-1 following our surface chemistry approach. XPS and antibody staining showed defined gradients of outwardly oriented SDF-1? active sites. This test platform can be especially valuable for investigators interested in studying the influence of surface-immobilized factors on cell behavior and may also be used as a cell migration enabling platform for testing the effects of various diffusible agents.

SUBMITTER: Vernekar VN 

PROVIDER: S-EPMC4259856 | biostudies-literature | 2014 Nov

REPOSITORIES: biostudies-literature

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Bi-ligand surfaces with oriented and patterned protein for real-time tracking of cell migration.

Vernekar Varadraj N VN   Wallace Charles S CS   Wu Mina M   Chao Joshua T JT   O'Connor Shannon K SK   Raleigh Aimee A   Liu Xiaji X   Haugh Jason M JM   Reichert William M WM  

Colloids and surfaces. B, Biointerfaces 20140919


A bioactive platform for the quantitative observation of cell migration is presented by (1) presenting migration factors in a well-defined manner on 2-D substrates, and (2) enabling continuous cell tracking. Well-defined substrate presentation is achieved by correctly orienting immobilized proteins (chemokines and cell adhesion molecules), such that the active site is accessible to cell surface receptors. A thiol-terminated self-assembled monolayer on a silica slide was used as a base substrate  ...[more]

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