Computerized analysis of tumor cells flowing in a parallel plate chamber to determine their adhesion stabilization lag time.
The importance of cell adhesion in a variety of physiological phenomena requires development of an understanding of the factors and molecular mechanisms underlying these behaviors. Cell adhesion is a multistep process involving primary receptor-ligand interactions followed by secondary events that may lead to the formation of focal contacts. Due to the lack of well-defined assays to study adhesion stabilization, little is known about this process, except that it may involve signaling events, receptor recruitment, and, as we have demonstrated, covalent peptide cross-linking by cell membrane-associated transglutaminase [Menter et al.: Cell Biophys. 18:123-143, 1992). To study the stabilization process we have developed a dynamic assay employing a parallel plate flow chamber coupled with video microscopy and digital image processing. Our studies utilize wheat germ agglutinin-selected human metastatic melanoma cell variants that exhibit differences in their experimental metastatic potential and expression of transglutaminase. Using this assay, quantifying cell-substrate stabilization was found to be quick, reliable, reproducible, and useful in evaluating agents that block this process.
Patton
J T
JT
Cox Laboratory for Biomedical Engineering, Rice University, Houston, TX 77251.
Menter
D G
DG
Benson
D M
DM
Nicolson
G L
GL
McIntire
L V
LV
D000368:Aged; D002448:Cell Adhesion / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002465:Cell Movement / Q000502:physiology; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008297:Male; D008545:Melanoma / Q000201:enzymology / Q000473:pathology* / Q000648:ultrastructure; D013997:Time Factors; D011503:Transglutaminases / Q000032:analysis / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured
D011503:Transglutaminases