Hollow-Fiber Assay for Ligand-Mediated Cell Adhesion
- Publication Type:
- Journal Article
- Citation:
- Cytometry Part A, 2004, 57 (1), pp. 39 - 44
- Issue Date:
- 2004-01-01
Closed Access
Filename | Description | Size | |||
---|---|---|---|---|---|
2008004473OK.pdf | 459.32 kB |
Copyright Clearance Process
- Recently Added
- In Progress
- Closed Access
This item is closed access and not available.
Background: The investigation of receptor-ligand interactions in the cellular context presents significant technical challenges, first, to immobilize the ligand in a manner that preserves functional properties and, second, to relate ligand properties to cell adhesion and other cellular processes. Methods: Ligand-mediated cell adhesion was characterized by the development of a cellulose hollow-fiber adhesion assay in which ligand (protein A) was immobilized onto the cellulose membrane as a recombinant fusion protein containing a cellulose-binding domain affinity tag. Modules containing single cellulose hollow fibers were connected to a micro-flow system for cell deposition and detachment with fluid shear stress. The cell adhesion process that occurred inside a segment of hollow fiber was observed in real time by using an inverted microscope equipped with a CCD camera and digital frame grabber. Image analysis software was developed to count cells and record digital images. Results: Cell adhesion strength was characterized by counting the number of cells that were detached by application of fluid shear stress with values that ranged from 2.3 to 185 dyne/cm2. The median shear stress of detachment of KG1a cells was directly related to the duration of membrane contact and the amount of immobilized monoclonal antibody (anti-CD34). Conclusions: The hollow-fiber assay provides a general method to determine functional properties of molecular domains that interact with cell surface receptors and markers. © 2003 Wiley-Liss, Inc.
Please use this identifier to cite or link to this item: